Enforced Bcl-2 expression overrides serum and feeder cell requirements for mouse embryonic stem cell self-renewal.
Leukemia inhibitory factor (LIF) is required, but not sufficient, for pluripotent mouse embryonic stem (ES) cell expansion in vitro in the absence of serum or a feeder cell layer, suggesting that additional signals are provided by serum or feeders that are necessary to support self-renewal. Here we show that transgenic ES cell lines expressing Bcl-2, an antiapoptotic protein, continue to self-renew in serum- and feeder-free conditions when supplemented with LIF; even in the absence of bone morphogenic proteins. Bcl-2-expressing clones sustain the characteristics of undifferentiated, pluripotent ES cells during long-term culture, and maintain their potential to differentiate into mature cell types. These results suggest that LIF and Bcl-2 overexpression are sufficient to expand these mouse pluripotent stem cells in vitro.
Yamane
Toshiyuki
T
Institute of Cancer and Stem Cell Biology and Medicine and Departments of Pathology and Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305, USA. yamanet@stanford.edu
Dylla
Scott J
SJ
Muijtjens
Manja
M
Weissman
Irving L
IL
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D001483:Base Sequence; D001769:Blood; D019485:Bone Morphogenetic Proteins / Q000378:metabolism; D002455:Cell Division; D002678:Chimera; D017931:DNA Primers; D004622:Embryo, Mammalian / Q000166:cytology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism* / Q000502:physiology; D013234:Stem Cells / Q000166:cytology*
D019485:Bone Morphogenetic Proteins; D017931:DNA Primers; D019253:Proto-Oncogene Proteins c-bcl-2
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