Document Detail


Simple and Efficient Derivation of Mouse Embryonic Stem Cell Lines Using Differentiation Inhibitors or Proliferation Stimulators.
MedLine Citation:
PMID:  21521035     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
The inhibition of endogenous differentiation-inducing signaling or the enhancement of growth capacity and viability of pre-implantation embryos, via 2i (PD0325901 and CHIR99021), dramatically improves the establishment of mouse embryonic stem (ES) cells. Using adrenocorticotropic hormone fragments 1-24 (ACTH 1-24), which enhances survival and/or proliferation of mouse ES (mES) cells, also increases the derivation of mES cells from single blastomeres significantly. The CHIR99021 pathway and the proposed ACTH pathway are likely different. Therefore, this study aimed to assess the synergetic effects of 2i and ACTH 1-24 on derivation of mES cells. Results in the present study demonstrate that germline transmitted mES cells could be derived efficiently from ICR and C57BL/6J at 0.5 to 4.5 days post-coitum denuded zygotes to blastocysts or isolated blastomeres of 2- to 8-cell embryos which cultured in 10-μL droplets with human foreskin fibroblast (Hs68) or STO feeders and in knockout serum replacement (KSR) ES cell medium containing 2i or ACTH 1-24. The overall success rates for C57BL/6J and ICR were 56.2% when cultured in 2i + ACTH 1-24, 26.6% in 2i, 6.7% in ACTH 1-24, and 4.8% in KSR ESC medium. These results imply that CHIR99021 and ACTH 1-24 are in synergistically enhancing the establishment of mES cells. The proposed protocol also demonstrate a highly efficient and reproducible method with a simple layout, and is easy to apply, and could be used as an alternative method for routinely establishing mES cell lines.
Authors:
Kun-Hsiung Lee; Chin-Kai Chuang; Shyh-Forng Guo; Ching-Fu Tu
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-4-26
Journal Detail:
Title:  Stem cells and development     Volume:  -     ISSN:  1557-8534     ISO Abbreviation:  -     Publication Date:  2011 Apr 
Date Detail:
Created Date:  2011-4-27     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101197107     Medline TA:  Stem Cells Dev     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Animal Technology Institute Taiwan, Division of Biotechnology, Chunan, Miaoli, Taiwan; khlee@mail.atit.org.tw.
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