山羊iPS细胞诱导及培养体系的优化

doi:10.11843/j.issn.0366-6964.2014.05.009

Abstract

Abstract:

In order to induced and cultured the goat iPS cells efficiently and continuous，the feeder cells and culture conditions were optimized in this study.MEF and GEF cells were inoculated at concentrations of 1×10⁵ mL^-1MEF,1×10⁵ mL^-1GEF and 5×10⁴ mL^-1MEF+5×10⁴mL^-1GEF as the feeder layers of goat iPS cells after they were treated with Mitomycin C within 3 passages，and cultured in DMEM+20%FBS,KnockoutDMEM+20%KSR，observed the different influence.The results showed that the induction efficiency and formation rates after digest of goat iPS cells in 5×10⁴ mL^-1MEF+5×10⁴ mL^-1GEF and Knockout DMEM+20% KSR were more higher than those cultured on the other 5 groups(P＜0.01).Growth states of goat iPS cells were observed in this group and undifferentiated phenotypes were detected，included expression of alkaline phosphatase，and dected the expression of Oct4,Sox2,Klf4,Nanog and cell marker Oct4,SSEA-1,Tra-1-60,Tra-1-81 by RT-PCR.In the condition of feeder layer free，goat iPS cells differentiation were observed in vitro. The results indicated that the acquisition and cultivate of goat iPS cells in 5×10⁴ mL^-1MEF+5×10⁴ mL^-1GEF and KnockoutDMEM+20% KSR were more easily than those cultured on the other 5 groups.This study established a foundation for further study on iPS cell in vitro studies，clinical trials，animal genome modification and goat ES cells line.

CLC Number:

S827
Q343

QIU Feng-long， ZUO Qi-sheng， LI Dong， LI Wei， CHEN Ting-feng， WEI Guang-hui， LI Bi-chun. Optimization of Induced and Cultivation System of Goat iPS Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(5): 740-749.

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Optimization of Induced and Cultivation System of Goat iPS Cells

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