Establishment and maintenance of embryonic stem-like cell lines from In Vitro produced bovine blastocysts = 체외수정 유래 소 배반포로부터 유사 배아 줄기 세포의 확립 및 유지

Abstract

This study was conducted to examine the establishment of bovine ES-like cells having pluripotency. The hatched blastocysts derived from culture of in vitro fertilized embryos for 10 to 12 days dissociated mechanically into ICM- and trophectoderm-rich clumps using needle, and cultured onto mitotically-inactivated MEF feeder layer. The primary colonies originated from ICM cells were detached mechanically 7 days after seeding and subsequent subculture was conducted at intervals of every 5 to 7 days. Two ES -like cell lines were established and maintained over 40 passages. Self-renewal of the established lines was confirmed by examining the alkaline phosphatase activity, stem cell-specific marker profiles including SSEA isotopes, Oct-4 and STAT3. Moreover, the established cell lines could produce anchorage-independent embryoid bodies (EBs) with gradual decrease of Oct-4 transcript level in time-dependent manner.