Culture in vitro and Cryopreservation of Shouguang Black Chicken Fibroblasts
[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 fibroblasts respectively, and the cell growth rates and the cryopreservation and recovery rates were compared. [Result] The primary chicken fibroblasts derived from enzymatic digestion grew faster than those derived from tissue explant culture and formed confluent monolayer after culture for 5 days. However, the passage fibroblasts derived from the two methods had similar growth rates, which could be cultured in confluence within 2 to 3 days. Homogeneous fibroblasts could be obtained after the mixture of fibroblasts and epithelial cells were treated by the combined use of typsin digestion and repeated attachment method for 3 or 4 subcultures. There were 75% -80% of cells survived after frozen and thawed. The separated and purified embryonic and skin fibroblasts showed normal and similar growth curves, both of which could be subcultured to the twelfth generation and still retain the normal chromosome number. [Conclusion] The feeder layer cells needed in ES cell line establishment could be obtained by culturing chicken fibroblasts using the tissue explant method and enzymatic digestion method. This study laid a foundation for the successful establishment of ES cell line.
SkinEmbryonicFibroblastsCulture in vitroCryopreservation
国家科技期刊平台
NSTL
万方数据
维普
