Basic fibroblast growth factor supports undifferentiated human embryonic stem cell growth without conditioned medium.
Previous studies have shown that prolonged propagation of undifferentiated human embryonic stem cells (hESCs) requires conditioned medium from mouse embryonic feeders (MEF-CM) as well as matrix components. Because hESCs express growth factor receptors, including those for basic fibroblast growth factor (bFGF), stem cell factor (SCF), and fetal liver tyrosine kinase-3 ligand (Flt3L), we evaluated these and other growth factors for their ability to maintain undifferentiated hESCs in the absence of conditioned medium. We found cultures maintained in bFGF alone or in combination with other factors showed characteristics similar to MEF-CM control cultures, including morphology, surface marker and transcription factor expression, telomerase activity, differentiation, and karyotypic stability. In contrast, cells in media containing Flt-3L, thrombopoietin, and SCF, individually or in combination, showed almost complete differentiation after 6 weeks in culture. These data demonstrate that hESCs can be maintained in nonconditioned medium using growth factors.
Xu
Chunhui
C
Geron Corporation, 230 Constitution Drive, Menlo Park, California 94025, USA. cxu@geron.com
Rosler
Elen
E
Jiang
Jianjie
J
Lebkowski
Jane S
JS
Gold
Joseph D
JD
O'Sullivan
Chris
C
Delavan-Boorsma
Karen
K
Mok
Michael
M
Bronstein
Adrienne
A
Carpenter
Melissa K
MK
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D000954:Antigens, Surface; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology*; D004815:Epidermal Growth Factor / Q000235:genetics; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005434:Flow Cytometry; D058851:GPI-Linked Proteins; D015870:Gene Expression / Q000235:genetics; D006023:Glycoproteins / Q000378:metabolism; D006028:Glycosphingolipids / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007621:Karyotyping; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D009363:Neoplasm Proteins / Q000235:genetics; D050814:Octamer Transcription Factor-3; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011509:Proteoglycans; D055746:Stage-Specific Embryonic Antigens; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D013724:Teratoma / Q000473:pathology; D060245:Tetraspanin 29; D014157:Transcription Factors / Q000235:genetics
D015703:Antigens, CD; D000954:Antigens, Surface; C554869:CD9 protein, human; C554870:Cd9 protein, mouse; D017077:Culture Media, Conditioned; D004268:DNA-Binding Proteins; D058851:GPI-Linked Proteins; D006023:Glycoproteins; D006028:Glycosphingolipids; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C494174:Pou5f1 protein, mouse; D011509:Proteoglycans; D055746:Stage-Specific Embryonic Antigens; C070636:TDGF1 protein, human; C107064:TRA-1-60 antigen, human; D060245:Tetraspanin 29; D014157:Transcription Factors; C066584:stage-specific embryonic antigen-4; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor; D019098:Telomerase