A New Chemical Approach to the Efficient Generation of Mouse Embryonic Stem Cells
Here, we present a highly efficient and reproducible method for the establishment of mouse embryonic stem cells (mESCs) from embryonic day 3.5 (E3.5) whole blastocysts. This protocol involves the use of small molecules SB431542 and PD0325901, which inhibit transforming growth factor-β (TGF-β) and extracellular signal-regulated kinases (ERK1/2), respectively. This protocol is universal in the derivation of mESC lines from NMRI, C57BL/6, BALB/c, DBA/2, and FVB/N strains, which have previously been considered refractory or non-permissive for ESC establishment. The efficiency of mESC lines generation is 100%, regardless of genetic background.
Key wordsMouse Embryonic stem cells derivation Small molecule ERK TGF-β
This study was funded by grants provided from Royan Institute, the Iranian Council of Stem Cell Technology, and the Iran National Science Foundation (INSF).
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