Long-term self-renewal and directed differentiation of human embryonic stem cells in chemically defined conditions.
Chemically defined medium (CDM) conditions for controlling human embryonic stem cell (hESC) fate will not only facilitate the practical application of hESCs in research and therapy but also provide an excellent system for studying the molecular mechanisms underlying self-renewal and differentiation, without the multiple unknown and variable factors associated with feeder cells and serum. Here we report a simple CDM that supports efficient self-renewal of hESCs grown on a Matrigel-coated surface over multiple passages. Expanded hESCs under such conditions maintain expression of multiple hESC-specific markers, retain the characteristic hESC morphology, possess a normal karyotype in vitro, as well as develop teratomas in vivo. Additionally, several growth factors were found to selectively induce monolayer differentiation of hESC cultures toward neural, definitive endoderm/pancreatic and early cardiac muscle cells, respectively, in our CDM conditions. Therefore, this CDM condition provides a basic platform for further characterization of hESC self-renewal and directed differentiation, as well as the development of novel therapies.
Yao
Shuyuan
S
Department of Chemistry, The Scripps Research Institute, SR202, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Chen
Shuibing
S
Clark
Julie
J
Hao
Ergeng
E
Beattie
Gillian M
GM
Hayek
Alberto
A
Ding
Sheng
S
D028341:Activins / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D019485:Bone Morphogenetic Proteins / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry*; D004622:Embryo, Mammalian / Q000033:anatomy & histology*; D004707:Endoderm / Q000166:cytology; D006801:Humans; D028322:Inhibin-beta Subunits / Q000378:metabolism; D007621:Karyotyping; D051379:Mice; D008819:Mice, Nude; D009206:Myocardium / Q000166:cytology; D009474:Neurons / Q000166:cytology / Q000502:physiology; D010179:Pancreas / Q000166:cytology; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D000099176:Noggin Protein
D028341:Activins; D015415:Biomarkers; D019485:Bone Morphogenetic Proteins; D002352:Carrier Proteins; D003470:Culture Media; D028322:Inhibin-beta Subunits; D000099176:Noggin Protein; C422405:activin A