Culture in vitro and cryopreservation of Shouguang black chicken fibroblasts
2008
Wang Juan | Yu Yuan | Wang Yuesi
[Objective] The aim of this study was to establish the in vitro culture system of chicken fibroblasts. [Method] Tissue explant method and enzymatic digestion method were used to separate and culture chicken skin fibroblasts respectively. The rate of cell growth, cryopreservation and recovery were compared. [Result] The primary chicken fibroblasts prepared by enzymatic digestion grew faster and converged together to form monolayer on 5 d post preparation; the passage cells prepared by these 2 methods grew at similar speed and formed monolayer within 2C3 d; homogeneous fibroblasts could be obtained by trypsin digestion and repeated attachment for 3C4 passages; there were 75%C80% of cells survived after cryopreservation and recovery; the growth curves of embryonic fibroblasts and skin fibroblasts were all normal and the two kind of cells still retained the normal number of chromosomes even at the twelfth passage. [Conclusion] The feeder layer cells needed for establishing ES cell lines could be obtained by culturing chicken fibroblasts through both tissue explant method and enzymatic digestion method. This study provided a basis for the successful establishment of ES cell lines.
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