We compared the characteristics of the method establishing embryonic stem cell lines from five different @entity19 strains using the medium containing 70% @entity35 heart cell-conditioned medium (RH-CM) as @entity2828 cell culture medium, using the primary @entity19 embryo fibroblast as feeder cells, and using the digestive enzyme buffer containing 1% chicken serum and "the series digestive method". We first reported new @entity2828 cell lines established from the outbred strain @entity19 KM and ICR using the improved method in our lab and the ratio of establishment of @entity2828 cell lines from KM and ICR strain @entity19 is up to 12% and 42.1% respectively. Compared with routine method of establishing @entity2828 cell lines, the improved method made distinct differences, increasing the ratio of @entity2828 cell line's establishment of 129/ter @entity19 from 11.8% to 33.3%, that of C57BL/6J @entity19 from 3.7% to 13.3%, that of BALB/c @entity19 from 2.9% to 19.4%. We tested the appropriate dispersing occasion, that is proliferating period of the ICM, affected the formation of @entity2828 clones and the ratios of @entity2828 cell lines established. It was shown that the most appropriate dispersed occasion for the ICM of 129/ter, C57BL/6J, BALB/c, KM and ICR @entity19 was 4-6 d, 3-3.5 d, 4 d, 4-5 d, 4-5 d after ICM proliferation respectively. At the same time, the effects of the concentration of digestive enzyme buffer were discussed. It was found that the @entity2828 cells from BALB/c @entity19 were sensitive to the high concentration of digestive enzyme buffer and the 0.05% Trypsin-0.008% @entity1098 is an ideal concentration for their establishment and maintenance. It was shown that 'the series dispersed method' was much better than 'the once dispersed method' on the aspect of dispersing the proliferating ICM and formation of @entity2828 clones. Compared with the routine @entity2828 cell culture medium containing mLIF, the RH-CM not only remarkably inhibited the differentiation of @entity19 @entity2828 cells and maintained their diploid karyotype, but also promoted the attachment and growth of @entity2828 cells. This improved method of establishment and culture of @entity2828 cell lines effectively maintained a series of their characteristics of pluripotent embryonic stem cells.

Comparison of the method establishing embryonic stem cell lines from five different XXXX strains.

@entity19 :: ('10090', 'Species') :: ['mice', 'mouse', 'murine'] @entity2828 :: ('MESH:D012512', 'Disease') :: ['ES'] @entity1098 :: ('MESH:D004492', 'Chemical') :: ['EDTA'] @entity35 :: ('10116', 'Species') :: ['rat']
@entity19 :: (10090,Species) :: ['mouse']
