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Serum replacement with a growth factor-free synthetic substance in culture medium contributes to effective establishment of mouse embryonic stem cells of various origins.

Abstract：

OBJECTIVE:To evaluate whether serum replacement with growth factor-free synthetic substances contributed to the effective establishment of embryonic stem (ES) cells. DESIGN:Randomized, prospective model study. SETTING:Gamete and stem cell biotechnology laboratory at Seoul National University in Korea. ANIMAL(S):F1 (C57BL6 x DBA2) mice. INTERVENTION(S):Blastocysts of different origins were cultured in serum-replaced media. MAIN OUTCOME MEASURE(S):Embryonic stem cell establishment. RESULT(S):Eight batches of ES cells were established from colony-forming inner cell mass cells after the replacement of fetal bovine serum (FBS) with synthetic knockout serum replacement (KSR) in mkDMEM. The established cells were positive for ES cell markers and formed both embryoid bodies in vitro and teratomas in vivo, but the established cell batches and control (transformed) ES cells responded differently to the culture media. Higher levels of cell viability were detected after the replacement with the 75:25 FBS-KSR mixture than with any other mixtures, and a gradual decrease in viability was detected as the KSR volume ratio was increased. The 75:25 FBS-KSR mixture-containing medium supported ES cell establishment of outbred ICR, F1, and F2 of C57BL6/DBA2; F1 parthenogenetic and ES cell-complemented tetraploid blastocysts; and single ES-cell cultures. CONCLUSION(S):A serum-replaced medium could be used for effective ES-cell establishment of various origins.

journal_name

Fertil Steril

journal_title

Fertility and sterility

authors

Lee ST,Oh SW,Kim DY,Han JY,Moon SY,Lim JM

doi

10.1016/j.fertnstert.2006.01.056

subject

Has Abstract

pub_date

2006-10-01 00:00:00

pages

1137-45

issue

4 Suppl

eissn

0015-0282

issn

1556-5653

pii

S0015-0282(06)01078-8

journal_volume

86

pub_type

杂志文章

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