Isolation and Cultivation of Embryonic Stem Cell Lines Derived from Pre-implantation Mouse Embryos of Four Different Strains

Abstract: To increase the efficiency of establishing mouse embryonic stem (ES) cell lines derived from various mouse strains. B6D2F1 (C57BL/6×DBA/2)， 129/SV×DBA/2， C57BL/6， BALB/C mouse were used in this report， blastocysts were obtained from 3.5 dpc (days post-coition) super ovulated mouse， or harvested morula from 2.5 dpc super ovulated mouse， cultured to blastocysts stage in vitro. Blastocysts transferred into ES culture medium with feeder cells， after 4? days of culture， inner cell mass (ICM) were picked up and transferred to the new culture dish with feeder cells， dissociate ICM by 0.05% trypsin after overnight culture， ES cells were passaged every 3? days. ES cell lines were established with obserration of phase contrast microscope and identified by karyotype， AKP staining， differentiation ability， SSEA-1， SSEA-4， TRA-1-60， TRA-1-81 immunostaining. 10 ES cell lines were established， each cell line exhibited typical mouse ES cell characteristics. 10 ES cell lines derived from B6D2F1 (C57BL/6×DBA/2)， 129/SV×DBA/2， C57BL/6， BALB/C mouse strains were established. Dissociating of ICM after overnight culture might be beneficial for establishment of mouse ES cell lines.

CSTR: 32200.14.cjcb.2007.04.0026