Induced Pluripotent Stem Cells (iPSCs) for research and therapy: induction of hepatic differentiation in iPSCs and evaluation of their quality as a model of in vivo development in the context of coagulation

.............................................................................................................................. 2 DECLARATION ....................................................................................................................... 3 ACKNOWLEDGMENTS ......................................................................................................... 4 TABLE OF CONTENTS ........................................................................................................... 5 LIST OF FIGURES ................................................................................................................... 8 LIST OF TABLES ................................................................................................................... 11 LIST OF SUPPLEMENTARY FIGURES .............................................................................. 12 LIST OF SUPPLEMENTARY TABLES ................................................................................ 13 ABBREVIATIONS ................................................................................................................. 14 Chapter 1 Literature review ................................................................................................ 17 1.1 Stem cells .................................................................................................................. 17 1.2 iPSC reprogramming ................................................................................................. 22 1.3 Applications of iPSC ................................................................................................. 31 1.4 Questions and challenges of the iPSC field .............................................................. 35 Chapter 2 Materials and methods ....................................................................................... 41 2.1 Cell culture ................................................................................................................ 41 2.2 Cloning ...................................................................................................................... 43 2.3 Reprogramming ......................................................................................................... 44 2.4 Pluripotency .............................................................................................................. 46 2.5 RNA isolation, RT-PCR and qPCR .......................................................................... 47 2.6 Phenotype analysis by flow cytometry...................................................................... 48 2.7 Immunostaining ......................................................................................................... 49 2.8 Hepatic differentiation............................................................................................... 49 2.9 Periodic Acid-Schiff (PAS) Stain for Glycogen ....................................................... 50