饲养层密度对兔类ES细胞分离培养的影响

doi:10.11843/j.issn.0366-6964.2013.07.009

Abstract

Abstract:

To explore the effects of different densities of feeder layer cells on isolation and culturing of rabbit ES-like cells and select the better culture system. Rabbit blastulas were cultured at different densities(10×10³, 20×10³, and 40×10³·cm^-2) of mouse embryonic fibroblast cell (MEF) as feeder layer, rabbit ES-like cell adherence and growth behavior were observed and express pluripotency markers were detected, such as AKP and Oct-4. The results showed that the attachment rate of blastulas and ICM formation rate at a density of 20×103·cm-2 feeder layer were higher than those at 40×10³·cm^-2 (82.5% vs 80.0%; 67.5% vs 65.7%, respectively), but the two groups were not significantly different (P>0.05). While stem cells cultured at a density of 20×10³·cm^-2 feeder layer were significantly higher than at 10×10³·cm^-2 feeder layer (82.5% vs 55.6%; 67.5% vs 44.4%, respectively) (P<0.05). The cell clone rate of F1、F2 generation of rabbit ES-like cells at a density of 20×10³·cm^-2 feeder layer was significantly higher than other two groups (P<0.05), and the stem cells obtained were up to the 20th passages and expression of AKP and Oct-4 were positive. In conclusion, the MEF feeder layer at a density of 20×10³·cm^-2 may be better suitable for rabbit ES-like cells culture in vitro.

CLC Number:

S829.1
Q813

LI Xiao-jia, ZHANG Jun-tao, XU Qiu-qin, WANG Ge, CAO Shu-jun, WANG Xin-zhuang, ZHANG Zhi-ping. Effect of Densities of Feeder Layers for Isolation and Culturing of Rabbit ES-like Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(7): 1051-1056.

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Effect of Densities of Feeder Layers for Isolation and Culturing of Rabbit ES-like Cells

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