The use of SC1 (Pluripotin) to support mESC self-renewal in the absence of LIF.
Mouse embryonic stem (ES) cells are conventionally cultured with Leukemia Inhibitory Factor (LIF) to maintain self-renewal.(1) However, LIF is expensive and activation of the LIF/JAK/STAT3 pathway is not absolutely required to maintain the self-renewal state.(2) The SC1 small molecule may be an economical alternative to LIF. SC1 functions through dual inhibition of Ras-GAP and ERK1.(3) Illustration of its mechanism of action makes it a useful tool to study the fundamental molecular mechanism of self-renewal. Here we demonstrate the procedure for culturing mouse ES cells in the presence of SC1 and show that they are able to maintain self-renewal in the absence of LIF. Cells cultured with SC1 showed similar morphology compared to cells maintained with LIF. Both exhibited typical mouse ES morphology after five passages. Expression of typical pluripotency markers (Oct4, Sox2, Nanog, and SSEA1) was observed after five passages in the presence of SC1. Furthermore, SC1 caused no overt toxicity on mouse ES cells.
Xiong
Wen
W
Research and Development, Stemgent. wen.xiong@stemgent.com
Gao
Yan
Y
Cheng
Xun
X
Martin
Charles
C
Wu
Dongmei
D
Yao
Shuyuan
S
Kim
Min-Ju
MJ
Liu
Yang
Y
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D003470:Culture Media; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects*; D036341:Intercellular Signaling Peptides and Proteins; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D051379:Mice; D010455:Peptides / Q000494:pharmacology*
D003470:Culture Media; D036341:Intercellular Signaling Peptides and Proteins; D053605:Leukemia Inhibitory Factor; D010455:Peptides; C076687:CS1 peptide