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1st ISSCCT

ABSTRACT BOOK


1st ISSCCT

ADDRESS: IUSCSC Istanbul University Fatih / ISTANBUL www.istanbulstemcell.com By 1st ISSCCT Organizing Committee Editor: Mücahid Erdoğan Cover design: Özgür Bölükbaşı


CONTRIBUTORS 1st International Students' Stem Cell Congress of Turkey HONORARY COMMITTEE Honorary President Prof. Dr. Osman İlhan Supervisor of Istanbul University Students’ Club of Stem Cell Prof. Dr. Tülay İrez Congress President Prof. Dr. Tülay İrez Special Thanks to... Prof. Dr. Teoman Soysal Prof. Dr. Selma Yılmazer Prof. Dr. Serap Erdem Kuruca Doç. Dr. Ahmet Tulga Ulus Prof. Dr. Osman İlhan Mr. Naim Kadoglou Prof. Dr. Francesco Dazzi Tamer Önder, PhD Asst. Prof. Halime Kenar

Matthias Lutolf, PhD Ömer H. Yılmaz, MD, PhD Anthony Gavalas, PhD Associate Professor Prof. Dr. Oktay Kadayıfçı Prof. Dr. Mehmet Ali Özcan Prof. Dr. Deniz Sargın Prof. Dr. Melek Öztürk Prof. Dr. Ayhan Bilir


CONTRIBUTORS CONTRIBUTORS Dialogue Atabay İlaç Onkim Municipality of Şişli Gümüşpınar MedSanTek Kurukahveci Mehmed Efendi Mahdumları Eti Çaykur Municipality of Istanbul IETT


1st ISSCCT ORGANIZING COMMITTEE Honorary President Prof. Dr. Osman İlhan

President Prof. Dr. Tülay İrez

Vice-presidents Burak Mergen, Tolga Aydın

Secretary General Kerem Fidan

Scientific Coordinator Betül Aksoy

Sponsor Relations Gamze Bildik, Aslı Semerci, İlkay Başak Uysal

Social Program Ece Ilgazlı, Hazal Haytural, Hazel Topçu

Public Relations Abdullah Keleş, Erkan Özergin, Elçin Ergez


1st ISSCCT ORGANIZING COMMITTEE

Treasurer Mücahid Erdoğan

IT Manager Yusufcan Koşansu

Press Cemre Kandaz, Tuğçe Ayık

Graphics & Design Özgür Bölükbaşı

Scientific Team Elif Saraçoğlu, Esra Hanedar

Senanur Şanlı

Simultaneous Translation Abdülkadir Baziki

Transportation Fırat Tevetoğlu, Rasimcan Meral

Foyer Area Dorukcan Alkan, Elif Gökçen Sazak Onur Erçelik


1st ISSCCT ORGANIZING COMMITTEE

Media Relations Bahar Engin

Internal Relations Murathan Türker

External Relations Ozan Ünlü

TURKSTEM ADMINISTRATIONAL BOARD Supervisor Assoc. Prof. Dr. Ahmet Tulga Ulus

President Tolga Aydın

Vice-President Burak Mergen

Secretary General Barış Ağar

Internal Relations Murathan Türker


1st ISSCCT ORGANIZING COMMITTEE External Relations Ozan Ünlü

Scientific Coordinator Betül Aksoy

Communication Sevda Aygun

Media Relations Bahar Engin

Treasurer Korcan Emre Gültekin


CONTENTS

Prefaces President of the 1st ISSCCT

Prof. Dr. Tülay İrez

1

Scientific Program

2

Oral Presentation Abstracts

6

1st Session – Introduction to Stem Cells 2nd Session – Neural Stem Cells 3rd Session – Cancer Stem Cells 4th Session – Tissue Engineering and Stem Cells 5th Session – Stem Cell and Surgery Applications 6th Session – Hematopoietic, Embryonic Stem Cells and iPS Cells

8 11 14 18 21 23

Poster Presentation Abstract

28

Awards and Jury Istanbul Medical Faculty Map Social Program Useful Information Sponsors Photo from 1st Stem Cell Student Symposium

31 32 33 34 35 36


PREFACE BY THE PRESIDENT OF 1st ISSCCT

PROF. DR. TÜLAY İREZ

Dear Colleagues, We are honored to welcome you to the 1st International Students’ Congress of Stem Cell. After new advancements on specific scientefic fields, stem cells have been fascinating subjects for laboratory researches and also promising tools for therapeutic applications since their discovery. Therefore, organizing congresses about Stem Cell’s becomes even more important because it allows us to share our novel informations and creates a social environment for participants to get know each other. The journey about Stem Cell has been started with our communication within our university but also with the members of TURK-STEM(Turkish Student’s Council of Stem Cell). TURK-STEM has been established with the very aim of providing a workplace for the medical students who are a candidate to become future’s scientists. On May 2011 being established with the leadership of Ankara, Hacettepe, Istanbul, Ege, Kayseri-Erciyes, Gata, Yeditepe, Celal Bayar universities. Also we would like to thank “Cellular Therapy and Regenerative Medicine Association” for their help about creating a platform for students who are willing to study stem cell. In Istanbul University, IUSCSC (Istanbul University Student’s Club of Stem Cell) gathered together in March 2011 and established formally in 1 June 2011 and my students honored me as their advisor. Our very aim is to provide vital information to everyone who is interested in this area and also provide them the essential laboratory opportunities to open their pathway with a multidisciplinary approach. In another words, we are willing to make an awareness about stem cell starting from our university ranging to other universities and citizens in Turkey. Our club’s members organized theoretical lectures every week and backed up these informations with laboratory courses since they have gathered in together. Also they have participated various congresses and been in internship programmes in Turkey and abroad. TURKSTEM has decided to initiate a series of students’ congresses about stem cell to be a first in the world and once again has shown how this council is a pioneer in it’s own field. With great efforts of our IUSCSC members and students whom take part in TURKSTEM administrative board we would all like to thank you for participating our congress which is the first of stem cell student congress in the world. We hope you enjoy the science, the spirit of the meeting and the setting at the oldest university in the Istanbul. Yours sincerely, Prof. Dr. Tülay İREZ President of 1st ISSCCT

1


SCIENTIFIC PROGRAM

1st International Students' Stem Cell Congress of Turkey in Istanbul Organized by IUSCSC & TURKSTEM

Scientific Program

2


SCIENTIFIC PROGRAM

13 May 2012 – 1st ISSCCT 08.00 – 09.00

On Desk Registration

09.00 – 09.40

Opening Ceremony

09.40 – 10.00

Coffee Break

10.00 – 11.30

1st Session: ‘‘Introduction to Stem Cells” Chairperson: Prof. Dr. Osman İlhan

10.00 – 10.45

Today, Past and The Future of Stem Cells

Prof. Dr. Francesco Dazzi, Keynote Lecturer

10.45 – 11.30

Philosophical Approach to Stem Cells

Oktay Kadayıfçı, MD Professor

11.30 – 11.40

Coffee Break

11.40 – 12.20

2nd Session: ‘‘Neural Stem Cell’’

Imperial College, London, UK Çukurova University, Turkey

Chairperson: Prof. Dr. Tülay İrez 11.40 – 12.20

Hox gene directed specification of diverse neural progenitors from pluripotent stem cells

12.30 – 13.30

13.30 – 14.30

Anthony Gavalas, PhD Associate Professor

Academy of Athens, Greece

Lunch Break

3rd Session: “Cancer Stem Cell’’ Chairperson: Prof. Dr. Serap Erdem Kuruca

13.30 – 14.10 14.10 – 14.30 14.30 – 14.50 14.50 – 15.50

mTORC1 in the Paneth cell niche couples intestinal stem cell function to calorie intake Effects of New Iron(III) and Nickel(II) Chelates of Smethyl-thiosemicarbazones on Cancer Stem Cells in K562 Cell Line Mode Separation and identification of cancer stem cell from leukemia cell lines by different ficoll density fractionation

Ömer H. Yılmaz, MD, PhD

MGH, USA

Erkan Özergin

Istanbul University

Abdullah Keleş

Istanbul University

4th Session: “Tissue Engineering and Stem Cell’’ Chairperson: Asst. Prof. Halime Kenar

14.50 – 15.30

Engineering artificial stem cell microenvironments

Matthias Lutolf, PhD Assistant Professor

EPFL, Switzerland

15.30 – 15.50

Human Endometrial Stem Cells a new source for Liver Regeneration

Sarah Dahmardeh

Tehran University of Medical Sciences, Iran

15.50 – 16.00

Coffee Break

3


SCIENTIFIC PROGRAM 16.00 – 16.40 16.00 – 16.40 16.40 – 18.00

5th Session: “Stem Cell and Surgery Applications” Chairperson: Tolga Aydın Stem Cell Applications in Cardiovascular Surgery

Ahmet Tulga Ulus, MD Associate Professor

TYIEAH, Ankara, Turkey

6th Session: ‘‘Hematopoietic, Embryonic Stem Cells and iPS Cells’’ Chairperson: Prof. Dr. Mehmet Ali Özcan; Prof. Dr. Deniz Sargın

16.40 – 17.20

Chromatin modifying enzymes as modulators of reprogramming

Tamer Önder, PhD

17.20 – 17.40

Erythroid differentiation of CD34+ hematopoietic progenitors is regulated by glucose and glutamine transporters

Leal Oburoğlu

17.40 – 18.00

Transplantation of autologous CD34+ G-CSF-mobilised peripheral mononuclear cells improves diabetic foot ulcers healing

Leila Mohammadzadeh

18.00 – 18.15

Poster Presentations

18.15 – 18.30

Closing Ceremony

4

Children’s Hospital Boston, USA Montpellier 2 University, France Tehran University of Medical Science, Iran


HEPATOLOGY

ORAL PRESENTATION ABSTRACTS

6


INTRODUCTION TO STEM CELLS

SESSION 1 INTRODUCTION TO STEM CELLS

SUNDAY MAY 13TH 10.00 – 11.30 CHAIRPERSON: Prof. Dr. Osman İlhan

8


INTRODUCTION TO STEM CELLS Prof. Dr. Francesco Dazzi About The Author

Professor Dazzi holds the position of Chair in Stem Cell Biology and Head of Stem Cell Biology at the Kennedy Institute of Rheumatology, Imperial College London. He is also academic leader of the stem cell transplantation programme there. His research interests are focused on the role of mesenchymal stem cells in normal and pathological conditions. He is searching into the factors which affect stem cell recruitment and function and developing various strategies to use these cells for regenerative medicine in degenerative and inflammatory diseases and to exploit their immunosuppressive properties in transplantation and autoimmunity. He has strong collaborative links with the Departments of Pathology (Prof Terence Cook) and Renal Medicine (Prof Charles Pusey) at the Hammersmith and with University College at the Royal Free Hospital (Prof Hans Stauss and Prof Stephen Mackinnon). Professor Dazzi publishes extensively in the area of mesenchymal stem cell immunoregulation and has publications in esteemed journals such as Blood, Stem Cells, Transplantation and Journal of Immunology.

Selected Publications: IlariaMarigo; Francesco Dazzi. The immunomodulatory properties of mesenchymal stem cells. Seminars in immunopathology 2011;33(6):593-602. Francesco Dazzi; Mauro Krampera. Mesenchymal stem cells and autoimmune diseases. Best practice & research. Clinical haematology 2011;24(1):49-57. Muzlifah A Haniffa; Matthew P Collin; Christopher D Buckley; Francesco Dazzi. Mesenchymal stem cells: the fibroblasts' new clothes? Haematologica 2009;94(2):258-63. Ingo MĂźller; Stefania Lymperi; Francesco Dazzi. Mesenchymal stem cell therapy for degenerative inflammatory disorders. Current opinion in organ transplantation 2008;13(6):639-44. Francesco Dazzi; Federica M Marelli-Berg. Mesenchymal stem cells for graft-versus-host disease: close encounters with T cells. European journal of immunology 2008;38(6):1479-82.

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13th

ORAL PRESENTATIONS –1st

Today, Past and the Future of Stem Cells


INTRODUCTION TO STEM CELLS Philosophical Approach to Stem Cells Oktay Kadayıfçı, MD Professor Half a cell comes from a mother and another from a father; sums up to form a single cell. The knowledge carried within that cell is such that with an appropriate habitat provided, the cell becomes a factory; producing life out of the raw material that is fed to it. The end product of the very fine line of production of the cell is replication; reorganization of products leads to two cells in the beginning, and then to a cluster of many cells by the continuum of this procedure. Then by a rather ambiguous, but certainly existent center of command, each cell is ordered to differentiate. This is only the beginning of a more complex mechanism that leads to the formation of tissues, organs and eventually organisms. In other words, every tissue and organ forming a living being stems from a single cell. This not only takes place in human, but in all living creatures. A more sophisticated self seems to exist within us, running beyond our conscious apprehension that happens to be far superior to our mind’s intellect. The same mechanism that produces new cells from others also facilitates the begetting of new organisms from former ones, heals wounds, combats harmful invaders to the body and destroys. In short, every single cell in our body possesses the knowledge and capability to form new tissues and organs from stem. Nevertheless, we already keep this process running in the background without us even knowing it. With scientific progress, we slowly started to bring into light how we are doing it. As we keep learning more and more, what has already been done in sheep might be possible on ourselves. If we can form an entire individual from a single cell, that means we should also be able to form individual organs of that being. After all, in case there are malfunctioning organs, we might be able to produce new ones and use in transplantation. With even further progress in our technologies, it might even be possible to do this by induction from outside, yielding surgery unnecessary. The first cell that has been fused from mother and father is a cell capable of transforming into every other type of cell. We may take any cell from a person’s body and insert some critical genes to make it imitate a cell that has newly been fused by mother and father. This cell we can then use in the same way as nature does to produce the organ or tissue we want. This is the logic behind stem cell today. Although we are yet in the crawling phase, we can already treat diseases like Leukemia, Cerebral Palsy, Type 1 Diabetes etc. Production of mouse liver and mouse bladder has been achieved. It is believed that in the near future there will be great progress in medicine with advances in stem cell, nano-technology, gene therapy and robotics; leading to higher quality of life in terms of health. About The Author He was born in Rize in 1973. He graduated from Kabataş Erkek Lisesi and Robert College in Istanbul. In 1967, he graduated from Istanbul Faculty of Medicine and he accomplished in the graduate student exam and in 1968 he became specialized student in Istanbul Faculty of Medicine’s Department of Obstetrics and Gynecology. In 1972 he became assistant doctor. After working for 2 months in this department he went to the Samsun Medical School for military service in 1972. He has been worked in Charing Cross Medical School for 2 years and during this time he was the director of “placental perfusion system and searching about the techniques of material exchange in the ongoing life of the placenta” research. In 1980 he received his Phd with the research about “placental perfusion system and searching about the techniques of material exchange in the ongoing life of the placenta”. In 1989 he received his professor title. Between the time in 1998 – 2009 he was the head of the Çukurova University’ Faculty of Medicine Department of Obstetrics and Gynecology. He is the manager responsible for the ONKIM Stem Cell Technologies and Cord Blood Bank since 2010.

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SESSION 2 NEURAL STEM CELLS

SUNDAY MAY 13TH 11.40 – 12.20 CHAIRPERSON: Prof. Dr. Tülay İrez

11

13th

ORAL PRESENTATIONS –2nd

NEURAL STEM CELLS


NEURAL STEM CELLS Hox gene directed specification of diverse neural progenitors from pluripotent stem cells Mina Gouti1, James Briscoe2 and Anthony Gavalas1 Timely induction of a single Hox gene (Hoxb1) in embryonic stem (ES) cell derived neural stem (NS) cells directed the generation of region specific neural progenitors. Dorsal morphogenetic signals resulted in strong up-regulation of neural crest (NC) genes selectively in Hoxb1+ NS cells implying the participation of Hox genes in NC induction. We addressed this by expressing Hoxb1 and other anterior Hox genes in the caudal neural tube of the developing chick embryo. Hoxb1 rapidly induced Snail2 and Msx1/2 and mobilized the entire NC specification program including epithelial to mesenchymal transition. These effects were dependent upon BMP signaling and optimal levels of Notch signaling. Other anterior Hox genes, such as Hoxa2, Hoxa1 and Hoxb2, but not the posterior gene Hoxb4 had a similar but weaker capacity. We extended this approach using different Hox genes to induce distinct NS cell fates corresponding to cervical (Hoxc6) and thoracic (Hoxc9) levels. We found that Hox genes modulate the interpretation of the ventral shh signaling and identified conditions resulting in enriched motor neuron progenitors (MNPs) corresponding to hindbrain, cervical and thoracic levels. To uncover specific signaling requirements for the specification and survival of MNPs we are analyzing the receptor tyrosine kinase (RTK) and nuclear receptor (NR) profiles of these cells. The implications of these findings in the design of efficient conversion methods of human stem cells into motor neurons for the study and treatment of motor neuron degenerative disorders will be discussed.

About The Author Anthony Gavalas received his Ph.D. from Purdue University (Indiana, U.S.) in 1994 in Molecular Biology and Biochemistry on the structure and regulation of genes encoding enzymes of the avian purine biosynthesis pathway. He pursued his postdoctoral studies as an EMBO and Marie Curie fellow in the labs of Pierre Chambon (I.G.B.M.C., Strasbourg) and Robb Krumlauf (N.I.M.R., London) on the role of Hox genes and retinoic acid in patterning the vertebrate hindbrain. In 2002 he joined the MRC Centre for Developmental Neurobiology as a Welcome Trust Career Development Fellow, the faculty of BRFAA in 2003 as a Researcher C' / Assistant Professor and the faculty of BRFAA in 2011 as a Researcher B’/Associate Professor. The main interests of his lab are (a) the directed differentiation of embryonic stem cells (ESCs) towards the neural and pancreatic endoderm lineages and (b) elucidation of cell fate specification mechanisms in the developing neural tube and pancreas of vertebrates. Selected Publications: Serafimidis, S. Heximer, D. Beis and A. Gavalas. GPCR signaling and S1P play a phylogenetically conserved role in endocrine pancreas morphogenesis (2011). Mol. Cell. Biol. 31, (5702-11). M. Bami, V. Episkopou, A. Gavalas and M. Gouti. Directed neural differentiation of mouse embryonic stem cells is a sensitive system for the identification of novel Hox gene effectors (2011). PlosOne, 6, e20197.

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CANCER STEM CELLS

SESSION 3 CANCER STEM CELLS

SUNDAY MAY 13TH 13.30 – 14.50 CHAIRPERSON: Prof. Dr. Serap Erdem Kuruca

14


mTORC1 in the Paneth cell niche couples intestinal stem cell function to calorie intake

Authors

Ömer H. Yılmaz, MD, PhD

How adult tissue stem and niche cells respond to the nutritional state of an organism is not well understood. Here, we find that Paneth cells, a key constituent of the mammalian intestinal stem cell (ISC) niche, augment stem cell function in response to calorie restriction (CR). CR acts by reducing mTOR complex 1 (mTORC1) signaling in Paneth cells, and the ISC-enhancing effects of CR can be mimicked by rapamycin. Calorie intake regulates mTORC1 in Paneth cells, but not ISCs, and forced mTORC1 activation in Paneth cells during CR abolishes their effects on ISCs. Finally, increased expression in Paneth cells of bone stromal antigen 1 (Bst-1), an ectoenzyme that produces the paracrine factor cyclic ADP ribose (cADPR), mediates the effects of CR and rapamycin on ISC function. Our findings establish that mTORC1 non-cell autonomously regulates stem cell self-renewal, and highlight a significant role of the mammalian intestinal niche in coupling stem cell function to organismal physiology.

About The Author Selected Publications: Levi BP, Yilmaz OH, Duester G, Morrison SJ. Aldehyde dehydrogenase 1a1 is dispensable for stem cell function in the mouse hematopoietic and nervous systems. Blood. 2009 Feb 19; 113(8):167080. Kiel MJ, Yilmaz OH, Morrison SJ. CD150- cells are transiently reconstituting multipotent progenitors with little or no stem cell activity. Blood. 2008 Apr 15; 111(8):4413-4; author reply 4414-5. Yilmaz OH, Morrison SJ. The PI-3kinase pathway in hematopoietic stem cells and leukemiainitiating cells: a mechanistic difference between normal and cancer stem cells. Blood Cells Mol Dis. 2008 Jul-Aug; 41(1):73-6. Yilmaz OH, Valdez R, Theisen BK, Guo W, Ferguson DO, Wu H, Morrison SJ. Pten dependence distinguishes haematopoietic stem cells from leukaemia-initiating cells. Nature. 2006 May 25; 441(7092):475-82.

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13th

Title

ORAL PRESENTATIONS –3rd

CANCER STEM CELLS


CANCER STEM CELLS Title

Effects of New Iron(III) and Nickel(II) Chelates of Smethyl-thiosemicarbazones on Cancer Stem Cells in K562 Cell Line Mode

Authors

Erkan Özergin(1), İzlem Yıldız(1) , Ali Aydoğan(2), Serap Erdem-Kuruca(3), Sema Bilgiç(4), Tülay Bal(5), Bahri Ülküseven(5)

Institutions Of The Authors

(1)

Cerrahpaşa Medical Faculty-English Program, Istanbul University, Istanbul, Turkey (2)

Cerrahpaşa Medical Faculty-Turkish Program, Istanbul University, Istanbul, Turkey (3)

Physiology Department, Istanbul Medical Faculty, Istanbul University, Istanbul, Turkey (4)

Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey

(5)

Department of Chemistry, Istanbul University, Istanbul, Turkey

Introduction

Cancer is a stem cell disorder and arose from normal stem cells. The conventional therapy modalities that target rapidly dividing cells rather than cancer stem cells (CSCs) have limited success in the cancer. CSCs have been shown to have different sensitivity to chemotherapeutic agents. Although the idea of the therapies focused on CSCs look more effective, they are not easy because CSCs are relatively quiescent and resistant compared to other cancer cells. Recently, thiosemicarbazones have drawn great interests for their high potential biological activity, especially their anticancer activity. However, there isn’t any report about their cytotoxic effects on CSCs. Therefore, in this study we tried to evaluate the antileukemic properties of metal complexes of new thiosemicarbazones synthesized by our team and their potential power to inhibit CSCs or stem-like cells.

Methods

Firstly, the iron (III) and nickel (II) chelates of some methoxy and dietylaminosal-substituted N1,N4-diarylidene-S-methylthio semicarbazones were synthesized. K562 chronic myeloid leukemia cells were cultured with these chelates in different concentrations. Cytotoxicity experiments were done by MTT [(3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide] assay. The concentration of the compounds which provides 50% inhibition of cell growth (IC50) were calculated from dose-response curve. Secondly, K562 cells are cultured with effective chelates in IC50. After three days viable cells were incubated with Rhodamine 123 (a fluorescent dye) and dye uptake of cells was determined with flow cytometry.

Results

Mean age in patients and controls was 11.44± 3.07 and 10±5.09 (p=0 .27) respectively. Tp-e had statistically significant difference between two groups (0.15± 0.07, 0.11± 0.04; p=0.003). ROC curve analysis showed that Tp-e larger than 0.14 has sensitivity of 47.4 and specificity of 85.7 and positive likely hood ratio of 3.32 for detection of patients with syncope.

Conclusion

Mean age in patients and controls was 11.44± 3.07 and 10±5.09 (p=0 .27) respectively. Tp-e had statistically significant difference between two groups (0.15± 0.07, 0.11± 0.04; p=0.003). ROC curve analysis showed that Tp-e larger than 0.14 has sensitivity of 47.4 and specificity of 85.7 and positive likely hood ratio of 3.32 for detection of patients with syncope.

Keywords

thiosemicarbazones, K562 cells, cytotoxicity, cancer stem cell, MTT assay

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Separation and identification of cancer stem cell from leukemia cell lines by different ficoll density fractionation

Authors

Abdullah KELEŞ, Gamze BİLDİK, Pelin ERSAN, Sema BİLGİÇ, Serap KURUCA

Institutions Of The Authors

Istanbul Medical Faculty, Istanbul University, Istanbul, Turkey

Introduction

Nowadays, the cancer may be considered as a cancer stem cell disorder rather than that of rapidly growing cells. Although the origin of the cancer stem cells is yet to be defined, the concept of the cancer stem cells may allow new treatment options in the possible cure of the cancer and new areas of research in carcinogenesis. However, further research is required to identify and separate the cancer stem cells in various cancers from normal stem cells and other cancer cells. Thus, development of new stem cell-based therapies is to be possible eliminating the residual disease and recurrence.

Methods

In this study, we purposed to separate cancer stem cells from leukemia cell lines. We eliminated sensitive clones treating with vincristine in low concentrations from K562 and HL60 cells along 3-4 weeks. Later, surviving cells were fractionated by different ficoll gradients (1070, 1067, 1055). Each gradient were counted by cell counter whereby cell size and determined to rhodamine 123 uptake by flow cytometry.

Results

Flow cytometry results show that there is no any Rhodamine 123 uptake difference in K562 cell fractions. But we recognize two different HL60 cell populations in HL60 fraction 3. There were two different Rhodamine 123 uptake cell populations. 46% of these cells had low Rhodamine uptake ability and 51% of these cells had high Rhodamine uptake ability. This difference shows us only the HL-60 gradient 3 cells have cancer stem cells’ characteristic. When we analyzed the cell size and cell number for each gradients, we show that in every gradient, most of the cells’ size was about 8-12 micrometer (70%) and other cells have 3-6 micrometer (30%) cell size.

Conclusion

In this preliminary study about cancer stem cell purification and identification, we show that low Rhodamine 123 uptake is important to show exist of the resistant cell. With more researches this study can be a method to test of the treatment.

Keywords

Cancer stem cell, Rhodamine 123, Vincristine

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13th

Title

ORAL PRESENTATIONS –3rd

CANCER STEM CELLS


TISSUE ENGINEERING AND STEM CELLS

SESSION 4

SUNDAY MAY 13TH 14.50 – 15.50 CHAIRPERSON: Assist. Prof. Dr. Halime Kenar; Assist. Prof. Dr. Matthias Lutolf, PhD

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Matthias P. Lutolf Proper tissue maintenance and regeneration relies on intricate spatial and temporal control of biochemical and biophysical microenvironmental cues, instructing stem cells to acquire particular fates, for example remaining quiescent or undergoing self-renewal or differentiation divisions. Despite rapid progress in the identification of relevant niche proteins and signaling pathways using powerful in vivo models, to date, many adult stem cell populations cannot be efficiently cultured in vitro without rapidly differentiating. To address this challenge, we have been exploring a combination between biomaterials engineering and microfabrication to generate powerful artificial microenvironments to study and manipulate stem cell fate in culture. These platforms allow key biochemical characteristics of stem cell niches to be mimicked and the physiological complexity deconstructed into a smaller, experimentally amenable number of distinct signaling interactions. The systematic deconstruction of a stem cell niche may serve as a broadly applicable paradigm for defining and reconstructing artificial niches to accelerate the transition of stem cell biology to the clinic. About the Author

Matthias Lutolf is head of the Laboratory of Stem Cell Bioengineering at EPF Lausanne, Switzerland. He is trained in Materials Engineering at ETH Zurich where he also carried out his Ph.D. studies on the development of a novel class of biologically responsive synthetic materials for tissue engineering (awarded with ETH medal in 2004). In 2005, Lutolf joined the Baxter Laboratory in Stem Cell Biology at the Stanford University School of Medicine to work on stem cells. He was awarded a Leukemia and Lymphoma Society fellowship (2006) for his research on ‘niche regulation of hematopoietic stem cells’. In 2007 Lutolf won a prestigious European Young Investigator (EURYI) award to start up his independent research. Research in the Lutolf lab is at the interface of stem cell biology and biomolecular engineering to gain fundamental insight on cell fate regulation by niches. Using a set of innovative bioengineering strategies, the Lutolf Laboratory is working on reconstructing the critical and specialized tissue-specific microenvironments, termed ‘niches’, which control the function of adult stem cells in our body. These well-defined in vitro niches are used to decipher adult stem cell regulation and may spawn new strategies for stem cell-based therapies. Selected Publications: Lienemann, P.S., Lutolf, M.P. and Ehrbar, M. (2012) Biomimetic hydrogels for controlled biomolecule delivery to augment bone regeneration. Advanced drug delivery reviews. Vannini, N., Roch, A., Naveiras, O., Griffa, A., Kobel, S. and Lutolf, M.P. (2012) Identification of in vitro HSC fate regulators by differential lipid raft clustering. Cell cycle (Georgetown, Tex.), 11(8). Lutolf, M.P. (2012) Materials science: Cell environments programmed with light. Nature, 482(7386) pp. 477-478.

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13th

Engineering artificial stem cell microenvironments

ORAL PRESENTATIONS –4th

TISSUE ENGINEERING AND STEM CELLS


TISSUE ENGINEERING AND STEM CELLS Title

Human Endometrial Stem Cells A New Source for Liver Regeneration

Authors

Sarah Dahmardeh

Institutions Of The Authors

Tehran University of Medical Sciences, Iran

Introduction

The registered clinical application base on stem cell technology has been considered at least a close to 10 years due to some restrictions such as the lack of suitable cell source and the risk of teratoma formation. The current study aims to introduce human endometrial stem cells (hEnSCs) as adult stem cells with the easy access source and no immunological response, for alternative cell therapy.

Methods

The hEnSC were exposed to hepatojenic induction medium for 30 days and hepatic differentiation was assess via quantitative reverse transcription-polymerase chain reaction, immunofluresent staining for hepatic-specific genes and proteins, including cytokeratin 18, alphafetoprotein (AFP) and albumin (Alb) as well as functional tests for glycogen storage was performed.

Results

Cultured cells on hepatogenic medium differentiated into hepatocytelike cells and expressed hepatic–specific markers on day 30 of differentiation. Periodic Acid-Schiff Staining showed that hEnSCs were able to storage of glycogen.

Conclusion

In the presence study, the differentiated cells features were closely resembled human adult hepatocytes by in vitro testing. These cells will be provided possible sources of cells for liver repopulation.

Keywords

Human endometrial stem cells, Hepatocyte-like cells, Differentiation, Liver regeneration.

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SESSION 5 STEM CELL AND SURGERY APPLICATIONS

SUNDAY MAY 13TH 16.00 – 16.40 CHAIRPERSON: Tolga Aydın

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13th

ORAL PRESENTATIONS –5th

STEM CELL AND SURGERY APPLICATIONS


STEM CELL AND SURGERY APPLICATIONS Stem Cell Applications in Cardiovascular Surgery Ahmet Tulga Ulus, MD Associate Professor Cardiac cell therapy, used to restore myocardial segments which have lost their vitality, is a treatment strategy giving exciting experimental results and it's clinical trials are accelerating. Unlike revascularization with catheters, medical therapies, electrophysiological applications, surgical revascularisation and other contemporary therapies for ischemic myocardial dysfunction, cell therapy has the potential to regenerate the damaged tissue and thus resolve the scar tissue. Evidence gathered through clinical and experimental studies is promising for future clinical benefits. For the cell therapy to become a routine clinical application, there are crucial steps such as purification of the donor cells, understanding of the mechanisms demonstrating how and which cell types to be used, generation of new tissues and restoration of the functions in the implanted tissue. Attentive selection of patients who will benefit the most, adherence to ethical regulations and group work principals should be kept in mind.

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SESSION 6 HEMATOPOIETIC, EMBRYONIC STEM CELLS AND iPS CELLS SUNDAY MAY 13TH 16.40 – 18.00

CHAIRPERSON: Prof. Dr. Mehmet Ali Özcan; Prof. Dr. Deniz Sargın

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13th

ORAL PRESENTATIONS –6th

HEMATOPOIETIC, EMBRYONIC STEM CELLS AND iPSCs


HEMATOPOIETIC, EMBRYONIC STEM CELLS AND iPSCs Chromatin modifying enzymes as modulators of reprogramming Tamer Ă–nder, PhD Generation of induced pluripotent stem cells (iPSCs) by somatic cell reprogramming involves global epigenetic remodeling1. While several proteins are known to regulate chromatin marks associated with the distinct epigenetic states of cells before and after reprogramming2,3, the role of specific chromatin modifying enzymes in reprogramming remains to be determined. To address how chromatin-modifying proteins influence reprogramming, we used shRNAs to target genes in DNA and histone methylation pathways, and have identified positive and negative modulators of iPSC generation. While inhibition of the core components of the polycomb repressive complex 1 and 2, including the histone 3 lysine 27 methyltransferase Ezh2, reduced reprogramming efficiency, suppression of SUV39H1, YY1, and Dot1L enhanced reprogramming. Specifically, inhibition of the H3K79 histone methyltransferase Dot1L by shRNA or a small molecule accelerated reprogramming, significantly increased the yield of iPSC colonies, and substituted for Klf4 and c-Myc. Inhibition of Dot1L early in the reprogramming process is associated with a marked increase in two alternative factors, Nanog and Lin28, which play essential functional roles in the enhancement of reprogramming. Genome-wide analysis of H3K79me2 distribution revealed that fibroblast-specific genes associated with the epithelial to mesenchymal transition lose H3K79me2 in the initial phases of reprogramming. Dot1L inhibition facilitates the loss of this mark from genes that are fated to be repressed in the pluripotent state. These findings implicate specific chromatin-modifying enzymes as barriers to or facilitators of reprogramming, and demonstrate how modulation of chromatin-modifying enzymes can be exploited to more efficiently generate iPSCs with fewer exogenous transcription factors.

About The Author Selected Publications: Kuo TC, Chen CT, Baron D, Onder TT, Loewer S, Almeida S, Weismann CM, Xu P, Houghton JM, Gao FB, Daley GQ, Doxsey S. Midbody accumulation through evasion of autophagy contributes to cellular reprogramming and tumorigenicity. Nat Cell Biol. 2011 Oct; 13(10):1214-23. Onder TT, Daley GQ. microRNAs become macro players in somatic cell reprogramming. Genome Med. 2011; 3(6):40 Gupta PB, Onder TT, Jiang G, Tao K, Kuperwasser C, Weinberg RA, Lander ES. Identification of selective inhibitors of cancer stem cells by high-throughput screening. Cell. 2009 Aug 21; 138(4):645-59

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Erythroid differentiation progenitors is regulated transporters

Authors

Leal Oburoğlu, Peggy Merida, Cédric Mongellaz, João Mamede, Jawida Touhami, Jean-Luc Battini, Marc Sitbon, Sandrina Kinet and Naomi Taylor

Institutions Of The Authors

Institut de Génétique Moléculaire de Montpellier, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5535, F-34293 Montpellier Cedex 5, France

Introduction

The function of cytokines in HSC differentiation has been extensively studied during the past decade but it is only recently that the importance of nutrient transport and metabolism in this process has been recognized. It is known that glucose as well as glutamine are essential “fuels” that contribute to the bioenergetic and biosynthetic requirements of primary and transformed cells. However, the energy source(s) which are critical for the expansion and differentiation of hematopoietic progenitors, and the regulation and interplay between the molecules transporting these nutrients during HSC differentiation have not yet been elucidated.

Methods

CD34+ human progenitor cells, isolated from umbilical cord blood, were expanded ex vivo and differentiated towards the erythroid lineage in the presence of recombinant erythropoietin (EPO). Expression of the Glut1 glucose transporter and the SCL1A5/ASCT2 glutamine transporter was assessed by RT-qPCR and changes in cell surface expression were monitored using retroviral-derived ligands. Furthermore, the specific requirements for Glut1 and ASCT2 in CD34+ cell expansion/differentiation were analyzed by introduction of the appropriate shRNAs.

Results

We find that erythroid differentiation of CD34+ progenitors is associated with increased levels of Glut1 and ASCT2 transcripts and the surface expression of both transporters is significantly enhanced upon acquisition of the erythroid marker Glycophorin A. Furthermore, both transporters are required for the optimal expansion and differentiation of CD34+ progenitors. Notably though, these transporters assure critical and non-overlapping functions during erythropoiesis; introduction of specific shRNAs against either transporter conferred a dramatic selective disadvantage, with a 4-fold decrease in shRNA expressing cells over a 3 day differentiation period.

Conclusion

The data presented here establish key roles for the Glut1 and ASCT2 transporters in HSC differentiation to the erythroid lineage. As expression of both transporters are required for effective erythropoiesis, our data point to specific roles for glucose and glutamine in this process. The interplay of these two nutrients in HSC survival and differentiation will be discussed.

Keywords

Hematopoietic differentiation

stem

cells,

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of by

CD34+ hematopoietic glucose and glutamine

metabolism,

Glut1,

ASCT2,

erythroid

13th

Title

ORAL PRESENTATIONS –6th

HEMATOPOIETIC, EMBRYONIC STEM CELLS AND iPSCs


HEMATOPOIETIC, EMBRYONIC STEM CELLS AND iPSCs Title

Transplantation of autologous CD34+ G-CSF-mobilised peripheral mononuclear cells improves diabetic foot ulcers healing

Authors

Leila Mohammadzadeh, Mohajeri

Institutions Of The Authors

Tehran University of Medical Science, Iran

Introduction

Several clinical trials have been shown that stem cells derived from bone marrow and CD34+ peripheral blood mononuclear cells (PBMCs) are able to improve angiogenesis and wound healing within ischemic tissues; but their Results were controversial. In current clinical trial, the role of stem cells derived from PBMCs in recovery of diabetic foot ulcers was investigated experimentally.

Methods

Twenty-one eligible volunteers admitted in the clinics of diabetes (Shariati and Rasul-e Akram hospitals, Tehran University of Medical Sciences, Tehran, Iran) between December 2008 and December 2010 were randomly divided into transplanted (n=7) and control (n=14) groups. G-CSF-moblised PBMNCs were injected to seven patients and the results were compared with 14 patients in control group. After three month follow-up, transplanted group studied ischemia manifestations, including pain and neurological signs, ulcer healing rate and lower limb amputation rate.

Results

Aankle-brachial index (ABI) and feeling the pulse within ischemic tissues of transplanted group was significantly improved (P= 0.035 and P= 0.01, respectively). Fifty percent (7 patients) of control group faced a major amputation (above the ankle) compared to none in transplanted group (P= 0.047). A significant difference in reduction of pain was found between the two groups (P= 0.014). The safety of transplantation was also confirmed by observing no consequences, including infection and immunological rejection.

Conclusion

This study provided further evidence that autologous transplantation of G-CSF-moblised PBMNCs is able to induce angiogenesis and wound healing process in patients with diabetic foot and CLI.

Keywords

autologous peripheral blood stem cells; diabetic foot ulcer; severe ischaemia

Bagher

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Larijani2,

and

Mohammad-Reza


POSTER PRESENTATIONS POSTER PRESENTATIONS

POSTER PRESENTATIONS

SUNDAY MAY 13TH 18.15 – 18.30

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POSTER PRESENTATIONS Title

Effect of deforolimus and VEGF on angiogenesis in endometrial stem cells following three-dimentional culture

Authors

Jafar Ai1, Seyed Mohammad Tavangar1, Seyed Naser Ostad1, Jamshid Hajati1, Sarah Dahmardeh2, Somayeh Ebrahimi Barough2, Armin Ai3, Anita Abbasi2

Institutions Of The Authors

1

Department of Tissue Engineering and Cell Therapy, Faculty of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran, Iran 2

Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran 3

Department of Biology, Faculty of Sciences, University of Tarbiat Moallem, Tehran, Iran Introduction

Endometriosis is the presence of endometrial tissue outside of the uterine cavity and is the most common gynecologic disorder in women of reproductive age. To determine the inhibitory effect of a deforolimus on angiogenesis in a three-dimensional (3-D) culture of human endometrial stem cells (hEnCs) in vitro. Angiogenesis is an important mechanism in the pathogenesis of endometriosis, and deforolimus has been shown to have anti-angiogenic activity.

Methods

This was an in vitro study of human endometrial stem cells in 3-D culture of fibrin matrix. The samples obtained from fundus of the uterine cavity were minced, stromal cells isolated and placed in a 3-D fibrin matrix culture system for angiogenesis with VEGF and inhibit angiogenesis by deforolimus. Finally these cells were prepared for flowcytometric analysis by CD31 antibodies.

Results

By 3-4 weeks, in cells treated with VEGF, endothelial cell branching was observed and rudimentary capillary-like structures formed. In the presence of 5ÂľM of deforolimus, angiogenesis was reduced. All of the results confirmed by CD31 flow cytometry.

Conclusion

The deforolimus were shown mechanisms of angiogenesis.

Keywords

endometrial stem cells, endometrium, 3-D culture, deferolimus, VEGF

be

effective

in

inhibiting

the

POSTER PRESENTATIONS 13 MAY

29

to


POSTER PRESENTATIONS POSTER PRESENTATIONS Title

Isolation of stem cells from Wharton’s Jelly of the Human Umbilical Cord and study of their differentiation ability to lens fibre cells

Authors

Seyedeh Mahsa Khatami, Masoud Maleki, Saber Zahri, Kamaloddin Hamidi

Institutions Of The Authors

1

Introduction

Recently the use of stem cells in many areas including cell therapy was expanded. In this study, the differentiation capacity of human Wharton’s jelly stem cells to lens fiber cells investigated.

Methods

In this experimental study, human Wharton jelly stem cells isolated using explant method. Then isolated cells was induced to lens fiber cells by Bovine vitreous and studied the morphological changes and expression of four crystallin gene including αA , αB , βB1 and βB3 crystallin by RT-PCR.

Results

hWJSCs was induced with bovine vitreous by 1:1 and 1:3 proporations for 10 days. After 10 days, induced cells were extensive and parallel to each other. In addition, regarding the molecular studies, four αA , αB ,βB1 and βB3 crystallin genes expressions were studied. αA-crystallin gene at 1:1 and 1:3 proporation and βB1-crystallin gene at 1:3 proporation were not expressed.Conversely, αB-crystallin and βB3crystallin gene at the above mentioned proporations and βB1-crystallin one at 1:1 proportaion were expressed.

Conclusion

Isolated cells of human umbilical cord Wharton's Jelly are very good source of mesenchymal stem cells for most of the research especially in differentiation studies. hWJSCs can differentiate to different cells like lens fiber cells. Using vitreous as lens fiber cells inducer showed αB , βB1 and βB3 expression in hWJSCs.

Keywords

Umbilical cord, Crystallin, Wharton's jelly, fiber cells, Mesenchymal stem cells

University of Mohaghegh Ardabili, Ardabil, Iran

2

Department of Biology, Science & Research Branch, Tabriz, East Azerbaijan, Iran

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JURY & AWARDS AWARDS 300 Euros

Best oral presentation

JURY All oral presentations are going to be evaluated primarily by the chairperson of the session with an evaluation form. Slide format, overall presentation, originality of the research, usage of time, discussion and Q&A will also have an important role on the final decision. The presenter should master his subject and be well-prepared for scrutinizing questions from the jury and chairperson.

Chairmen Prof. Dr. Osman İlhan Prof. Dr. Tülay İrez Prof. Dr. Serap Erdem Kuruca Asst. Prof. Halime Kenar Tolga Aydın Prof. Dr. Mehmet Ali Özcan Prof. Dr. Deniz Sargın Jury Prof. Dr. Tülay İrez Assoc. Prof. Dr. Ahmet Tulga Ulus

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MAIN ENTRANCE

Istanbul Medical Faculty Map

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SOCIAL PROGRAM With a view of the Golden Horn: "Maxigala"

Maxigala under the Galata bridge, one of the surprises of Istanbul. During the dinner, watching the ferries, people and the sunset will be a completely different experience. Outdoor sitting atmosphere is great, you can enjoy it while you sipping your cold and hot drink.

We will meet at the great scenery with a unique history, eat delicious Turkish food and have fun a lot! While we enjoying our meal, we can also enjoy with a DJ performance. We'll dance to the sea,drink our beverages; this night will be an unforgettable for all of us!

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USEFUL INFORMATION METRO MAP 1 2 3 4 5

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Atatürk Airport (Havalimanı) Findikzade (Kaya Hotel) Eminonu Taksim Çapa-Şehremini (Istanbul Medical Faculty)

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PHOTO OF 1st NATIONAL SYMPOSIUM

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