Cornelia Kasper completed her Ph.D. 1998 from Leibniz University of Hannover (Germany) and her habilitation in 2007 at the Institute for Technical Chemistry at the Leibniz University of Hannover. She was appointed as full University Professor for "Biopharmaceutical Production and Technology" at University of Natural Resources and Life Science (BOKU) in Vienna (Austria) at the Department of Biotechnology. She has published more than 80 papers in reputed journals and several book chapters and is editor within the series "Advances in Biochemical Engineering and Biotechnology" (Springer) of several volumes covering actual areas in Tissue Engineering and Stem Cell Research. She is also reviewer for many distinguished journal within the field of biotechnology/bioprocess and bioreactor design and enableling technolgies for stem cell cultivation.

Introduction: Multipotent mesenchymal stromal cells (MSC) are being used with favorable success for tendon regeneration in large animal models as well as in equine patients. Tenogenic differentiation and subsequent matrix synthesis by the MSC was hypothesized to be one of the mechanisms of MSC-supported tendon healing. However, the tenogenic differentiation pathway is not yet completely understood. It is acknowledged that not only growth and transcription factors, but also the extracellular matrix or scaffold composition and mechanical stimulation play a crucial role. Therefore, our aim was to combine natural tendon scaffolds and mechanical stimulation in an in vitro bioreactor model as a basis for the investigation of MSC tenogenic differentiation.
Materials and Methods: To obtain scaffolds with natural matrix proteins in parallel alignment, large equine tendons were decellularized as described previously (Burk et al., 2013). 2 mm thick scaffolds were cut from the decellularized tendons and subjected to mechanical testing. Based on the results of the mechanical scaffold assessment, a cyclic strain bioreactor prototype was designed. The prototype was tested using equine adipose derived MSC which were cultured on the tendon scaffolds and subjected to cyclic strain stimulation (2% strain; 1 Hz; 15 min stimulation- 15 min relaxation- 30 min stimulation). Samples were assessed 24h later by LIVE/DEAD® staining, histology and real-time RT-PCR.

Mette Hemmingsen has completed her Ph.D. about culture and differentiation of cells in micro fluidic array systems and is currently doing postdoctoral studies in liver tissue engineering from The Technical University of Denmark, Department of Micro- and Nanotechnologies.

A huge shortage of organs for transplantation has motivated the research field of tissue engineering to develop bio-artificial liver tissue and even a whole liver. Due to limitations of primary hepatocytes regarding availability and maintenance of functionality, stem cells and especially human induced pluripotent stem cells (hIPS cells) are an attractive cell source for liver tissue engineering. Our approach for engineering liver tissue is to culture and differentiate hIPS cells in a 3D porous polymer scaffold built-in a perfusable bioreactor to ensure supply of oxygen and nutrients and removal of waste. To investigate the hepatic differentiation at flow conditions, IPS-derived definitive endoderm cells were seeded into the scaffold and differentiated according to a protocol developed by our collaborator Cellectis AB. Compared to conventional batch cultures, a similar hepatocyte-like cell morphology and gene expression of the hepatocyte nuclear transcription factor 4α was observed, whereas a decreased expression was for seen for the transcription factor CAR and the CYP enzymes CYP3A5 and CYP3A7. Furthermore, expression of albumin and α-fetoprotein was almost knocked down. However, expression of all markers was increased by the use of conditioned medium or medium with a two times lower concentration of the signalling factors than those optimized for conventional batch cultures. Thus, our results suggest that the flow conditions affect paracrine cell signalling necessary for liver differentiation and/or functionality, as well as the concentration of signalling factors in the differentiation medium has to be adapted to the different environment at flow with constant renewal of the culture medium.

Shaden Khalifa is a certified physician with bachelor degree in Medical and Surgical Sciences. Shaden has completed her Master Degree at biomedical Science from Uppsala University and her Ph.D. from Karolinska Institute, Stockholm, Sweden after 2 years of research visit to Keio University, Japan. Dr. Shaden has published 10 papers in reputed journals most of them are within the area of tissue engineering and regenerative medicine.

Electronic interfaces between living cells or tissues and conducting surfaces that serve to pass signals between the electronic devices and a living organism present an enormous scientific and technological challenge. Such bioelectronic interfaces are now widely used in medical devices including pace makers, cochlear implants and neurostimulatory implants. These devices have become indispensable in many situations and extended or dramatically improved the lives of millions of people. For example, worldwide cochlear implants have been performed in more than 100.000 persons and the current installed base of pacemakers is presently about 600.000 in the United States alone. Thus even modest improvements in these devices would quickly translate into improvements in human health and substantial advances would have tremendous impact. There is also a great deal of activity in other types of devices based on bioelectronics interfaces. Strategy to rescue hearing and auditory nerve was developed using micro-contact technique. Surface for nerve guidance was designed with favorable extracellular proteins to promote the outgrowth of the neurons. Micro-contact imprinting provided a versatile and useful technique for patterning the guidance surface. Imprinting generated a patterned surface in a controllable, predictable, and quantifiable manner. A range of events followed the patterning including alignment, polarity and directionality was reported and observed by morphological and microscopic description. The dynamic microenvironment that results from the synergistic combination of extracellular guidance cues and Schwann cells selectively instruct and direct the terminal extension of neurons into uni- or bi-polar fate. Applying new factors such as molecules, cells and surfaces provides unique possibilities to recruit spiral ganglion neurons into their regenerative ability. In summary, creating an environment that incorporates multiple molecular and cellular cues will offer exciting opportunities for elucidating the mechanisms behind nerve regeneration and highlight specific considerations for the future tissue engineering.

Chiara Gomiero completed her Degree in Industrial Biotechnology at University of Padua, Italy in 2010. Since then, she was a Temporary Research Assistant in Xeptagen SpA in Venice and she worked, for one year, in the diagnosis of new tumor biomarkers for the diagnosis and treatment of the main solid tumors of the adult. Then she won a bursary with Telethon at the Department of Biomedical Sciences of University of Padua where she studied three genetic diseases that affect striated muscles: type 2D Limb Girdle Muscolar Dystropy, Brody's disease and CPVT. Actually is a PhD Student in Dept. Comparative Biomedicine & Food Science at University of Padua and works with stem cells for the regeneration of muscle, tendon and epithelial pathologies. She presented abstracts/posters/talks in some conferences.

The polypeptide TAT is a small molecule essential for viral replication that possesses the unusual property of entering the cells from the extracellular milieu. Several experiments using recombinant GFP fused, either to TAT or to its basic domain, showed that the internalization process occurs through caveolar endocytosis. In order to produce tenocytes and myocytes from undifferentiated peripheral blood MSC, we present an innovative methodology based on the use of transcription factors Scleraxis and MyoD fused with the sequence TAT which promotes the cellular internalization without the use of viral vectors. MyoD and Scleraxis are two transcriptional factors that respectively promote myogenesis and tenogenesis. The nucleotide sequence encoding human MyoD was amplified from a human cDNA library while the nucleotide sequence encoding mouse Scleraxis gene was amplified from plasmid RCAS(B) mScx (Addgene). The amplified products were cloned in plasmid pTAT-Ngl to generate the fusion protein TAT-MyoD and TAT-Scleraxis containing the translocation domain of the HIV-1 proteinTAT (underlined)(MRGSHHHHHHGMARGYGRKKGRQRRR). The fusion proteins were then expressed and purified adapting standard recombinant techniques. In this study we used the undifferentiated MSC collected with a non-invasive withdrawal from peripheral blood (pbMSC) of horse. We have tested different incubation times to detect when the proteins fused with TAT were fully internalized in the nucleus of pbMSC and we have demonstrated that MyoD-TAT and Scleraxis-TAT protein can be found in the cell nucleus after few hours from the inoculation. Finally, we have evaluated the myogenic and tenogenic differentiation of MSC; for myogenic induction cells were induced by MyoD-TAT and co-cultured with mouse myoblast cell line C2C12 whereas, for tenogenic induction, cells were induced by Scleraxis-TAT with the addition of specific growth factorsas BMP-12 and FGF-2.The differentiation towards myogenic and tenogenic lineages of pbMSC were analysed using immunofluorescentspecific antibodies.

In the tissue regeneration field the use of scaffolds as support represents an important complementary tool to surgery in tendon and skin tissue engineering. Up to now natural and synthetic materials have been used as extracellular matrix (ECM) equivalent but none of them simultaneously offered biocompatibility, biofunctionality and mechanical performances. Collagen-made materials are the most promising for this purpose. Commercially available collagen is mainly of bovine origin, but in recent times alternative sources of collagen were found, including aquatic organisms. In the present work we used common marine invertebrates (echinoderms) as an innovative and low-cost source of native intact collagen fibrils to develop ECM equivalents. These animals possess peculiar connective tissues, called Mutable Collagenous Tissues (MCTs), which display striking passive mechanical properties. The idea we pursue is that MCT-derived collagenous matrices have the potential to be used for tissue engineering/regenerative medicine, especially for those applications where high mechanical performances are required (e.g. tendons).Native intact collagen fibrils were isolated from echinoderm connective tissues and used to prepare films and 3D scaffolds. In vitro biocompatibility and cytotoxicity tests were performed usingmesenchymal stem cells isolated from horse peripheral blood (PB-MSCs) and cultured for different time-points using the MCT-derived matrix as substrate; in vivotests were assessed by subcutaneous implantation of the MCT-derived matrix in rabbits.The evaluation of cell viability, inflammatory response and angiogenesiswere performed with histo-immunohistochemical and molecular methods. The obtained fibrillar matrices, which strictly mimicked the native ECM, were used as a support for the growth and proliferation of PB-MSCs; in vitro tests indicated that: MCT-derived matrices are not toxic for mammalian cells, showed standard (fibroblast-like) morphology and behaviour and, after an initial “adaptation” stage, were also able to proliferate actively. In vivo tests and post mortem histological analysis suggest encouraging results related to biocompatibility, although further investigations will be necessary.

Sylwia Koniusz has graduated from the Warsaw University of Life Sciences with master’s degree in biotechnology. She did the research for her bachelor’s degree at the Department of Cellular Engineering at the Cancer Centre in Warsaw and the research for her master’s degree at the NeuroRepair Department at the Mossakowski Medical Research Centre Polish Academy of Sciences in Warsaw. In October 2013 she has started at the same department the MMRC-KNOW Interdisciplinary Ph.D. Studies with the project „The role of bone marrow mesenchymal stem cells and microvesicles derived from these cells in CNS repair of brain ischemia disorders”.

Stem cells have been recognized as a potential tool to restore cells damaged by cerebral ischemic injury. Key functions such as the replacement of neural cells have been recently challenged by intrinsic bystander capacities of undifferentiated donor cells. One of opportunity for neurological disorder treatment is the transplantation of mesenchymal stem cells (MSCs) which have neuroprotective, neuroregenerative and anti-inflamatory properties. However, a comprehensive knowledge how transplanted MSCs exert their therapeutic achievements is still lacking. The aim of the project was to analyze the presence, distribution and quantity of human bone marrow mesenchymal stem cells (hBM-MSCs) transplanted into focal brain ischemic rats.The experiments were performed in adult male Wistar rats withbrain focal ischemiainduced with 1μl/50nmol ouabain(sodium-potassium pump inhibitor)injection into right stratium. Then 5x105 hBM-MSC (Lonza) stained with iron nanoparticles and rhodamine (Molday, BioPAL) were transplanted into internal carotid artery, 48 hours after brain insult. At 1, 3, 7 and 14 days rat brains were removed. Immunocytochemical analysis of human markers using different antibodies anti: CD44, STEM121and Ku80 were performed. The preliminary results showed that after intra-arterially injection of hBM-MSC, the donor cells were present in the ipsilateral rat hemisphere between cortical cortex and stratium near the ischemic lesion. The positive staining for Molday particles and human antigens were observed at 1, 2, 3 and 7 days after hBM-MSC transplantation. The further studies relating to the function of transplanted cells are in progress.

Haizea Iribar B.Sc. by Autonomous University of Barcelona (2010). The aim of my Ph.D. project is to elucidate the ontogeny, expansion and differentiation capacity of neural precursor cells in mouse and human dermis, with the ultimate purpose of facilitating their therapeutic use through generation of tissue engineered constructs and help clarify their possible contribution to carcinogenic processes or other pathologies.

Currently unpublished studies in our group show that human neural precursor cells (hNPCs) resident in the dermis can be traced back to Schwann and perivascular niches. In vivo and in vitro fate analyses showed that neural competence is restricted to Schwann cell lineage and that it correlates with high Sox2 levels (Perez-San Vicente et al., 2014). Dermal cells may have diverse embryonic origins, from Wnt1+ neural crest in the head to Myf5+ presomitic mesoderm in the dorsal trunk skin and lateral plate mesoderm in the ventral trunk dermis. Thus, we aimed to investigate the embryonic origin of NPC in the ventral dermis, also analyzing the Sox2 implication in neural competence of this cell population. For this, we carried out lineage tracing analyses using Myf5-Cre (dermomyotome), Cspg4-Cre (perivascular and glial cell marker), and Sox10-Cre (neural crest) transgenic lines crossed with EYFP reporter mice. FACS-based dermal cell isolation in those models followed by neurogenic differentiation showed a major contribution of Myf5+ cells to the dermis-derived neurogenic precursor cell population in ventral trunk. Correlation studies with endogenous Sox2 levels are underway.

Luiz Augusto U. Santos is Master in Science and Implantodontist at University of Sao Paulo. He works as Manager of Tissue Bank and Researcher of the Institute of Orthopedics and Traumatology, Hospital das Clínicas of the School of Medicine of the University of Sao Paulo. Over the years, he has studied the clinical use of bone allografts in orthopedic and dental surgeries and has published studies nationally and internationally. He is a member of the American Association of Tissue Bank (AATB) and the American Dental Association (ADA). He is also a member of the Group of the Regulatory Health Organizations related to the definition of legislation and standards.

Introduction: Bone loss has long been a challenge to dental surgeons who seek to reconstruct and rehabilitate patients with resorptions, especially in the maxillary and mandibular grafting. A range of biomaterial is available for use in dentistry, however the homologous graft, or allografts, has been increasingly used. This study explores four methods for processing the allografts (Group A: fresh allograft; Group B: Irradiated and Criopreserved bone allograft (- 80°C ); Group C: Freeze- Dried Bone Allograft; Group D: Demineralized Bone Allograft. It was initially performed a qualification testing (phase 1: sterility and cytotoxicity tests) and further an analysis of its osseointegration in an experimental trial in rabbits (phase 2: Optical Coherence Tomography (OCT), Cone Beam Tomography, Raman spectroscopy, histomorphometryand immunohistochemistry.
Objectives:
The aim of this study is to evaluate the bone allografts that were processed in four different ways:
A) Criopreserved allograft
B) Criopreserved - Irradiated allograft
C) Freeze-Dried allograft
D) Demineralized - Freeze-Dried allograft
Methods: To analyze the osseointegration, the areas of grafts were submitted to the exams of Optical coherence tomography (OCT), Cone Beam Tomography, Raman spectroscopy, histomorphometry and immunohistochemistry.

Neia completed B.Sc. in Biotechnology by University of Vic (2010) and Master degree in Biomedicine by University of Barcelona (2011). I have worked in different labs during my university studies and I have been involved in different scientific projects as an apprentice. At present I am working as a predoctoral student to develop an in vitro system for muscle engineering, with some of the results just published. The goal of my PhD project is to identify and characterize a myogenic cell resident in the adult skin of mice and humans and then generate a functional human muscle 3D-culture.

We have recently shown that bona fide, pulsating skeletal muscle myofibres may be generated from dermis-derived cells through recreation of 3D myogenic niche (Garcia-Parra et al., 2014). Interestingly, and after one month culture engineered muscle constructs showed progressive degradation of the myofibres concomitant with fatty infiltration, paralleling the natural course of muscular degeneration. However a critical point to translate these results to humans is to determine the origin and identity of myogenic precursor cells enriched within murine dermal cultures. Knowing that dermal and muscle cells share a common embryonic origin at the dermomyotomal stage, and taking into account that there might be different types of cells within adult skin presenting myogenic potential, our main objective was to identify and characterize the origin and identity of myogenic cells present in dermal cultures. To this end, we tested as working hypotheses the enrichment of (i) satellite cells from the dermal Panniculus carnosus (PC) muscle, (ii) dermomyotome-derived adult stem/precursor cells, (iii) perivascular cells, and (iv) neural crest-derived precursor cells. In order to trace the origin and identity of dermal myogenic cells, we took advantage of the following transgenic mice to perform lineage tracing experiments: (i) Pax3-GFP and Pax7CE (tracers of PC-derived satellite cells); (ii) Myf5-Cre (dermomyotome), (iii) Cspg4-Cre (perivascular and glial marker), and (iv) Sox10-Cre (neural crest). Cell tracing combined with FACS-based isolation and myogenic differentiation assays showed a major contribution of Myf5+ cells to the dermis-derived myogenic precursor cell subset, which was at least in part derived from PC satellite cells.

Chen was initially trained in Biochemistry and Molecular Biology during MS study in Huazhong Agricultural University, China. Since joining Key Laboratory for Regenerative Medicine, Ministry of Education, as a technical staff at Ji Nan University in 2008, he has been working on dissecting gene regulation mechanisms using cardiac microvascular endothelial cells as a model system. His current work is focus on the functional roles of miRNAs in regulating gene expression in ageing and myocardial infarction.

Background: Myocardial infarction (MI) was the leading cause of death in worldwide. MicroRNAs (miRNAs) regulate gene expression at the post-transcriptional level and are known to play essential roles in various aspects of biological processes, including cell viability, proliferation, development and differentiation. The purpose of this study was to investigate difference of miRNA profiles between infarct zone and border zone in post-MI remodeling using the second generation sequencing.
Results: We induced MI by ligation of the left coronary artery (LAD) in rat. The miRNA profiles between infarct zone and border zone were compared in 14 days after MI using deep small RNA sequencing, and followed by bioinformatics analysis. It was found that 25,603,683 and 26,050,631 clean reads were obtained in infarct zone and border zone respectively, and 234 conserved known miRNAs and 5 novel potential miRNA candidates were identified. The digital expression profiling based on TPM (Transcripts Per Million) analysis of conserved miRNAs show that a total of 22 miRNAs were identified the most highly detected miRNAs (>1000 TPM) in the infarct zone and 27 miRNAs in the border zone. A total of 40 conserved microRNAs were significantly differentially expressed between infarct zone and border zone. Comparing with miRNA expression in the infarct zone, 29 miRNAs were up-regulated and 11 miRNAs were down-regulated in the border zone. In addition, 7283 annotated mRNA transcripts of selected miRNA were identified as putative target genes using microRNA Target Filter tool from Ingenuity Pathway Analysis (IPA) (Ingenuity Systems). IPA pathway analysis suggested that the differentially expressed miRNAs are involved in many molecular and cellular functions, such as Cell-To-Cell Signaling and Interaction, Cellular Assembly and Organization, Cellular Function and Maintenance, Cellular Development, Cell Death and Survival and so on.
Conclusions: Our findings contribute to understanding the role of differentially expressed miRNAs between infarct zone and border zone, which show that miRNAs are dynamically regulated in different regions of the heart during post-MI remodeling.

Miriam C. Amores de Sousa is currently a PhD student at Department of Bioengineering of Instituto Superior Técnico (IST), in University of Lisbon (UL). Miriam thesis is focused on study the interaction between electrospun functional nanofiber matrices and neural stem cells, to evaluate cell fate envisaging applications in tissue engineering. Miriam graduated in Applied Chemistry (minor in Biotechnology) in 2007 and concluded the Master in Biotechnology in 2009, in Universidade Nova de Lisboa (UNL). Previously, as Research Assistant, Miriam worked on the characterization ofbiocompatible cellulose acetate membranes as potential drug delivery systems, focusing on the solid-state mobility properties.

Therapeutic strategies based on stem cell (SC) therapies require large cell numbers, howeverthese are found in low numbers in the human body. Therefore, there is a call for ex-vivo scaled-up processes in order toovercome the limited availability of SCs.When envisaging regeneration of neural tissue, cultivation of neural SC in alignednanofiberscaffolds is particularly attractive. Such substrates actually providephysical guidance with high surface-to-volume ratio, contributing to cell attachment andorientation, in structures mimicking the natural extracellular matrix (ECM) environment. Culture of SC within nanofiber scaffolds is usually performed in static systems, often neglecting the scalability required for the systematic production of tissue transplants, or tissue engineered platforms. Effective supplyof adequate factors (nutrients cytokines, growth factors, oxygen),cell interactions and shear forces are also critical. Therefore anovel scalable system for culture of stem cells combining cues from controlled hydrodynamic conditions withelectrospun scaffoldsfor guidance of human neural stem cells (hNSC) cultureis presented. In this work, the flow structure and wall shear stress was investigated by computational fluid dynamics. The system was also evaluated through the expansion of ahNSC population over 18 days, reaching a 3.5 fold increase in cell number, witha population that expanded uniformly along the nanofibers. These results are promising for scale up production of tissue constructs for regenerative medicine, drug testing or other biomedical applications.This work was co-financed by BIOREG (SOE3/P1/E750)-INTERREG IV-B SUDOE Program with ERDF funds;information on this project is included.

Anna Andrzejewska has completed her studies in Faculty of Biology at University of Warsaw at the age of 24. She obtained her master degree in biotechnology with specialization in molecular biology in 2012 and then started her PhD studies in Mossakowski Medical Research Centre Polish Academy of Sciences at NeuroRepair Department under supervision of Barbara Łukomska Professor of Immunology and Transplantology. She has already attended 5 conferences including 2 international conferences where she presented short communication of her results. She has published one paper and currently 2 more publications are in preparation.

Application of mesenchymal stromal cells (MSCs) is a new and promising approach in neurological disorders. The success of clinical application of MSCs relies upon the efficient recruitment of these cells into appropriate tissues. Systemic infusion of MSCs is preferred as a minimally invasive method of transplantation however cell migration from the vasculature into injured brain is inefficient. The aim of our study is to improve MSCs homing by genetic cell engineering leading to VLA-4 over expression. This is expected to cause extensive adhesion of MSCs to the endothelial cells assessed in this project using microfluidic devices. Materials and Methods:hBM-MSCs (Lonza) were selected for the study. Over expression of α4 subunit of VLA-4 integrinin hBM-MSCs was obtained by mRNA-ITGA4 based transfection. The naïve (non-transfected)and modified (mRNA-ITGA4 transfected) hBM-MSCswere stained with iron nanoparticles coupled with rhodamine (Molday, BioPAL) and used in microfluidic assay with microchannels coated with rat brain endothelial cell line (RBE-4). Results and Conclusions: During the in vitro observation naïve hMSCs interacted with rat brain endothelial cells. After the pressure-driven MSCs perfusion in microfluidic channels that can mimic blood flow in capillaries, the naïve cells were observed to roll, capture or arrest at the surface of endothelial cells layer. The efficiency of modified MSCs adhesion is in progress. The model of microfluidic assay is a useful tool which can be used to assess the adhesive properties of engineered MSCs before their intravenous or intra-arterial transplantation.

Michaela Hajkova is a Ph.D. Candidate at Charles University in Prague, Faculty of Science. She finished her master’s degree in Biology with the thesis: “The role of selected cell populations and molecules in inflammatory reaction and rejection of skin allograft”. She is working in the laboratory of immunoregulation where she is participating in research of therapeutic potential of MSCs and their ability to contribute to tissue repair after transplantation of mouse skin allograft.

Mesenchymal stem cells (MSCs) represent a population of multipotent stem cells with immunomodulatory, antiapoptotic and cytoprotective capabilities and thus hold a great promise for treatment of many inflammatory diseases and for use in a regenerative medicine. Numerous studies have shown that the administration of MSCs in combination with an immunosuppressive drug prolongs allograft survival in comparison with use of MSCs or the drug alone. However, the exact mechanism of such synergism has not yet been described. The aim of this study was to analyze the therapeutic effect of MSCs applied locally on polylactic acid (PLA) nanofiber scaffold with or without incorporated Cyclosporine A (CsA) in a mouse model of allogeneic skin transplantation. It was founded that treatment with MSCs and CsA significantly decreased the amount of graft-infiltrating macrophages and the production of nitric oxide. These changes were accompanied with considerable lower production of interferon-gamma as cytokine priming classically activated macrophages. Simultaneously, increased production of interleukin-10 by graft infiltrating macrophages and significant upregulation of CD206 expression as a marker of alternatively activated macrophages was observed. Our results indicate that application of MSC-seeded and CsA-loaded nanofiber scaffolds direct macrophage polarization towards development of alternatively activated macrophages. This phenotype switching may result in suppression of the local inflammatory reaction and takes part in healing process.

Miriam C. Amores de Sousa is currently a PhD student at Department of Bioengineering of Instituto Superior Técnico (IST), in University of Lisbon (UL). Miriam thesis is focused on study the interaction between electrospun functional nanofiber matrices and neural stem cells, to evaluate cell fate envisaging applications in tissue engineering. Miriam graduated in Applied Chemistry (minor in Biotechnology) in 2007 and concluded the Master in Biotechnology in 2009, in Universidade Nova de Lisboa (UNL). Previously, as Research Assistant, Miriam worked on the characterization ofbiocompatible cellulose acetate membranes as potential drug delivery systems, focusing on the solid-state mobility properties.

Therapeutic strategies based on stem cell (SC) therapies require large cell numbers, howeverthese are found in low numbers in the human body. Therefore, there is a call for ex-vivo scaled-up processes in order toovercome the limited availability of SCs.When envisaging regeneration of neural tissue, cultivation of neural SC in alignednanofiberscaffolds is particularly attractive. Such substrates actually providephysical guidance with high surface-to-volume ratio, contributing to cell attachment andorientation, in structures mimicking the natural extracellular matrix (ECM) environment. Culture of SC within nanofiber scaffolds is usually performed in static systems, often neglecting the scalability required for the systematic production of tissue transplants, or tissue engineered platforms. Effective supplyof adequate factors (nutrients cytokines, growth factors, oxygen),cell interactions and shear forces are also critical. Therefore anovel scalable system for culture of stem cells combining cues from controlled hydrodynamic conditions withelectrospun scaffoldsfor guidance of human neural stem cells (hNSC) cultureis presented. In this work, the flow structure and wall shear stress was investigated by computational fluid dynamics. The system was also evaluated through the expansion of ahNSC population over 18 days, reaching a 3.5 fold increase in cell number, witha population that expanded uniformly along the nanofibers. These results are promising for scale up production of tissue constructs for regenerative medicine, drug testing or other biomedical applications.This work was co-financed by BIOREG (SOE3/P1/E750)-INTERREG IV-B SUDOE Program with ERDF funds;information on this project is included.

Dossova Aigul in 1995 she graduated Karagandy State Medical University by the specialty “Medical care”. Since 1995 till 1997 – research assistant in Otolaryngology department. 1997 – 2000 – Doctoral student in the same department. In 2001 – defense the work on the topic: “Clinical and Pharma-kynetic argumentation of lymphatic acute otitis media and estimation its efficiency”. Since 2002 she has been working for JSC National Scientific Medical Research center on the position of leading research worker of the department of organ and cell transplantation. She is a developer of cellular therapy at diseases of otitis media and internal ear in Kazakhstan. She is an author of different works concerning indicated problem.

Introduction Treatment of traumatic perforations is an actual problem in ENT surgery. It is connected with its prevalence rate, especially in working age young people [1]. One of the classical methods of reconstruction of tympanic membrane after traumatic perforation is myringoplasty, which is attended with definite surgical difficulties [2]. One of the attempts of solving this question is using of cell transplantation, in particular also fetal fibroblasts, coming with apparent regenerative properties [3]. Today clinical experience of application of cell technologies in ENT practice is minimal, that is connected with poor research and practical materials, difficulties of fetal cells obtaining and it is attended with insufficiently worked-out methodical and methodological approaches of its application [4]. Purpose of study is effectiveness estimation of influence of fetal fibroblasts on processes of regeneration at traumatic perforation of tympanic membrane. Materials and approaches After experimental animal studies [5], within the framework of scientific agenda of the National Scientific Medical Research center «Innovative technologies in development of cell transplantations and reconstruction of functional activity of organs and tissues», on the basis of MOH of the RK order «On permission of using fetal cells transplantation in comprehensive treatment of chronic diseases» № 211 as of 22 April, 2009, permission of academic council (protocol №3 as of 19 February 2010) and Ethics Committee «NSMRC» (protocol №2 as of 22 January, 2010), the decision to use this method of treatment in ENT pathologies was made. On 20.04.2010 patient A., 25 years old, was hospitalized to ENT Department with posttraumatic perforation of tympanic membrane. After necessary examination patient was conducted transplantation of fetal fibroblasts. Culture of fetal fibroblasts was collected under the laboratory conditions of cell technologies according to developed by us method (innovative patent № 19899 Agency on innovations of the MOH of the RK) from the derma of 18-20 week fetus through cultivation in Iscov «Sigma» environment with addition of FBS 10% «HyClone». After 14 days of cultivation monolyer of fetal fibroblasts was received, which were washed out from the environmental components and inserted into collagenic matrix for further transplantation. Deepithelialization of the edge of ruptured hole was conducted after anesthesia of auditory passage skin, whereafter transplant from fetal fibroblasts on collagenic matrix in the amount of 2х106 cells in 0, 5 ml of physiological solution was placed on humid wound surface of tympanic membrane. Otoscopic and audiometric studies were conducted overtime for estimation of processes of reconstructive regeneration of tympanic membrane and auditory acuity after cell transplantation.

Matthias Graupp is a resident for otorhinolaryngology at the Medical University of Graz since 2011. He has completed his study of medicine at the age of 25 years at the Medical University of Graz. In 2012 he started his Ph.D studies with the project “Establishing a novel in-vitro model for vocal fold scarring”. He has published eight papers in reputed journals.

Introduction: An in vitro model that mimics vocal fold (VF) fibrogenesis is highly desirable but is not realized so far. Such a model would be of great value for rapid screening of antifibrotic compounds before going into larger expensive clinical trials and would create a real alternative to animal studies. The development of a fibrogenesis model has been hampered by several biological and technical obstacles. The biggest challenge is to guarantee a sufficient in vitro production of collagen and its incorporation into a pericellular matrix. Fibrogenic cells in monolayer culture do not lay down significant amounts of stable collagen in a useful time for testing antifibrotic compounds. Many of these obstacles might be overcome by implicating the principles of the excluded volume effect (EVE) and macromolecular crowding (MMC). Methods: VF fibroblasts from Sprague-Dawley rats were cultured either in standard medium, as well as in “crowded medium” by adding inert macromolecules (Ficoll®). Differentiation into myofibroblasts was achieved by administration of TGFβ-1 (transforming growth factor beta-1). Cells were cultured in standard medium/crowded medium, with or without administration of TGFβ-1. After five days culture medium and cell layer were collected separately and analyzed. Results: Collagen deposition increased significantly under crowded conditions in both culture medium and cell layer. Administration of TGFβ-1 resulted in a significant increase of collagen deposition compared to zero-control. Amounts of collagen in the cell layer were significantly higher under crowding conditions compared to administration of TGFβ-1 alone. Discussion: Crowding of the culture medium resulted in a significant increase of collagen deposition compared to standard medium in both culture medium and cell layer. In this specific environment collagen processing was enhanced, resulting in more favorable conditions for studying fibrogenesis compared to conventional cell culture medium. This can be the first step not only towards developing a rapid and robust in vitro model for testing of antifibrotic compounds, but also in the construction of ECM components.

Alessandro Bertolo has completed his Ph.D. at the age of 32 years old at University of Bern, Switzerland. His Ph.D. focused on the development of tissue engineering approaches based on mesenchymal stem cells application to treat intervertebral disc degeneration. He is currently involved in intervertebral disc studies both at in vitro and in vivo levels, as postdoctoral fellowship. Previously, he worked as fellow at S. Raffaele hospital, Milan, in the field on neural stem cells.

Mesenchymal stem cells (MSCs) are fundamental players in future cell-based therapies. These cells possess high proliferative ability, multilineage potential and immunomodulatory properties, all requisites essential for cell therapies. Nevertheless, intra-donor variability of MSCs is not fully characterized yet, leading to uncertain outcomes in the use of these cells. Here we propose a life imaging tool to characterize population of human MSCs derived from different donors, based on the in vitro mobility of the cells and to correlate results to the respective differentiation potential. Bone marrow human MSCs from 20 donors and at various in vitro passages (from P3 to P13) were evaluated. Track length of 70 cells per population were measured over a period of 24 hours, and distances were compared with the respective adipogenic, chondrogenic and osteogenic differentiation potential. We found that senescent populations were enriched with slow moving cells, compared with younger ones. Cells with larger cell body were moving less compared to smaller ones, while spindle shaped cells had an average speed. Our preliminary data showed that both fast cells and slow cells were characterized by low differentiation potential, while average moving cells were more effective in undergoing all three lineage differentiation. Heterogeneity in single cell motility within a population correlated with faster moving cells, while populations rich in slow moving cells tended to homogeneity (only senescent cells). In conclusion, in vitro cell mobility might be an useful tool to characterize MSC population and recognize prior to use the differentiation potential of MSCs.

Hepatocyte transplantation is a promising alternative to orthotopic liver transplantation for the treatment of liver diseases; however, the shortage of donor organs available to isolate good-functional quality cells is a major limitation. Neonatal livers may be a potential source alternative to adult liver to obtain good-performing hepatic cells for hepatocyte transplantation, which has not yet been explored in depth. High-yield preparations of viable hepatocytes were isolated from 1 to 23-day-old liver donors, cryopreserved and banked, and cell integrity and functional quality assessment were performed after thawing. Neonatal hepatocytes showed better post-thawing recovery if compared with adult hepatocytes, as shown by the viability values not differing significantly from freshly isolated cells, a higher expression of adhesion molecules(β1-integrin, β-catenin and e-cadherin), better attachment efficiency and cell survival, and a lower number of apoptotic cells. If compared to adult liver, the hepatocyte isolation procedure in neonatal livers also provides thawed cell suspensions with a higher proportion of hepatic progenitor cells (EpCAM+ staining), which could also participate in regeneration of liver parenchyma after transplantation. Additionally, when transplanted into the animal model of Crigler-Najjar disease (Gunn rat), an increased attachment and a long-term effect has been observed with newborn hepatocytes compared to adult ones. These results could imply important advantage to neonatal hepatocytes as a source of high quality cells to improve human hepatocyte transplantation applicability.

Minhao Li is a senior student from the Kunshan High School, Jiangsu, China. He has been working on a project of bone tissue engineering during his research internship under the supervision by Dr. Lee Chou, Professor of Biomaterials of Boston University.

Biomimetic design and 3--‐D printing of scaffold with heterogeneous internal geometry is essential for cell distribution, blood vessel in growth and biomaterial degradation in bone tissue engineering. This study was designed to demonstrate the heterogeneous pores and channels in 3--‐D printed scaffolds for bone tissue engineering. Scaffolds were prepared using ceramic particles through 3--‐D printing. Pores and connecting channels with diameters of 200 mm --‐ 500 mm were designed for facilitating cell seeding and cell distribution. Internal pores of 50 mm --‐ 200 mm were designed for bone regeneration. Nano--‐sized surface topography was designed for enhanced degradation of scaffold. The fabricated scaffolds were evaluated using scanning electronic microscopy. SEM of fabricated scaffolds revealed that 400 mm--‐ 500 mm inter--‐connecting channels crossed over the entire scaffold, that ~200 mm internal pores were scattered over the scaffold and connected to each other and to the inter--‐connecting channels, and that ~200 nm pores showed on the surfaces of inter--‐connecting channels and internal pores, which would play an important role in increasing the surface ratio of materials and facilitating material degradation. A heterogeneous profile of connecting channels and internal pores was evident in these 3--‐D printed biomimetic scaffolds. As a conclusion, the biomimetic design and fabrication of scaffolds for bone tissue engineering can be fulfilled by a 3--‐D printing process. Heterogeneous profiles of inter--‐connecting channels, internal pores, and nano--‐sized surface topography can be generated to provide a biomimetic environment suitable for bone tissue engineering.

Nelson Lizier holds a BS in Biotechnology, Masters of Science and PhD at the Laboratory of Genetics at the Butantan Institute and Department of Morphology and Genetics, Federal University of São Paulo. Has experience in the area of General Biology with an emphasis in cell and molecular biology research with stem cells: embryonic stem cells, germline stem cells, induced pluripotent stem cells (iPS) and mesenchymal stem cells. TOP 100 Scientists 2014, International Biographical Centre - Cambridge -England

Immature stem cells from dental pulp (IDPSC) are originated from neural crest and express embryonic, mesenchymal and epithelial stem cell markers in vitro. They appear to be less problematic being not ethically controversial and painful source, widely multipotent, not tumorigenic, which maintain their ‘‘stemness’’ for several serial passages. Because of short population doubling time, these cells can be up-scaled in large numbers. Due to these characteristics, they considered the most promising tool for regenerative medicine and tissue engineering. Aiming at establishing cell bank of IDPSC, here we compare IDPSC isolated at the very beginning (early) of dental pulp tissue culturing and after three months (late). The phenotype and proliferative potential and differentiation capacity were evaluated, when IDPSC were cultured in different media. Our data demonstrate that IDPSC can be easily isolated and successfully expanded in vitro, while maintain their undifferentiated status preserving their properties even after cryopreservation. Furthermore, dental pulp tissue can be maintained long time in culture providing unlimited source of IDPSC for future therapeutic applications. We also demonstrated that no significant changes in proliferative potential, stem cells markers expression profile and differentiation capacities in both, early and late isolated IDPSC populations were observed. These data can contribute significantly to the practical application of IDPSC in regenerative medicine. Therefore, this report describes that the hurdles of adult stem cells technologies have been overcome in the case of IDPSC. These stem cells present biotechnological product ready to go throughout stem cell-based therapy market.

Tania Limongi is research scientist in the Physical Science and Engineering Department of the King Abdullah University of Science and Technology (KAUST) in Saudi Arabia. She received in 1999 her Master Degree in Biological Sciences at the University of L’Aquila and her Ph.D. in Microsystem Engineering at the University of Rome Tor Vergata in 2004. She published more than 30 papers in reputed journals; her work is mainly addressed on the development of new tissue engineering approaches and on X-ray, TEM and Atomic Force microscopy application for the study of DNA/protein interactions and single molecule localization techniques.

Scaffold design and fabrication are very important subjects for biomaterial, tissue engineering and regenerative medicine research playing a unique role in tissue regeneration and repair. Polycaprolactone (PCL) is a very attractive bioresorbable polyester due to its high permeability, biodegradability and capacity to be blended with other biopolymers. By virtue of its ability to naturally degrade in tissues, PCL hold immense promise as new materials for implantable biomedical micro devices. This work focuses on the establishment of a microfabrication process integrating lithography and micromolding fabrication techniques for the realization of 3D microstructured PCL devices. Scaffold’s surface exhibits a combination in the patterned length scales; cylindrical pillars of 10 um height and 10 um diameter were arranged in a hexagonal lattice with periodicity of 30 um and theirs sidewalls were nano-sculptured with a regular pattern of grooves leading to a spatial modulation in the z direction. To demonstrate that these biocompatible pillared PCL substrates are suitable to a proper cell growth, NIH/3T3 mouse embryonic fibroblasts were seeded on microstructured substrates and their key adhesion parameters were evaluated. Scanning Electron Microscopy and immunofluorescence analysis were done to check cell survival, proliferation and adhesion; cells growing on the PCL substrates appeared healthy and formed a well-developed network in close contact with the micro and nano features of the pillared surface. Those highly flexible scaffolds could be a promising candidate solution for a wide range of tissue engineering and regenerative medicine applications.

Manarbek B. Askarov, MD, DMs (June 17, 2009), has the rank of professor (pathological physiology, transplantation and artificial organs). He is the head of the laboratory ofbone marrow stem cell biotechnology and transplantation in non-hematopoietic pathology at National Scientific Research Medical Center, Astana, Kazakhstan. He published more than 80 scientific papers (research articles, book chapters and abstracts and posters at various international conferences).

Introduction The progression of chronic heart failure is due to not only neurohormonal , but also immune mechanisms, which are implemented by the syndrome of chronic systemic inflammation . We believe that bone marrow stem cells by producing peptides, growth factors and cytokines that promote inflammation reducing inhibition activity of fibroplastic processes and stimulate reparative (replacement) regeneration of myocardial tissue . Purpose: To evaluate the safety and efficacy of systemic administration of bone marrow mononuclear cells (BMMC) in patients with severe left ventricular systolic dysfunction (EF < 40%) and dilatation syndrome. Research methods: We examined 27 patients ( 25 men and 2 women ) with marked dilatation syndrome and low left ventricular fraction (EF < 40%), aged 60 years. Of them with true dilated cardiomyopathy - 11 patients , ischemic cardiomyopathy- 14 , periportal cardiomyopathy in 2 women. Fence of bone marrow was performed in aseptic conditions of the operating unit from the iliac bone of the patient . Then using biotechnological method mononuclear cell fraction was allocated. Preliminary precultivating process plays an important role in therapy with autologous bone marrow cells to restore their bioregulatory activity. After cultivating for 72 hours , the suspension of BMMC administered systemically (intravenously). All patients received basic treatment with CHF ( ACE inhibitors , B- blockers, aldosterone antagonists , diuretics ) . Quality of life was determined by the " Minnesota questionnaire quality of life in patients with CHF (MLHFO)». Results were processed using the Student's test