寿光黑鸡成纤维细胞的体外培养与冷冻保存(英文)

[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 2-3 d;homogeneous fibroblasts could be obtained by trypsin digestion and repeated attachment for 3-4 passages;there were 75%-80% 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.

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 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 2-3 d; homogeneous fibroblasts could be obtained by trypsin digestion and repeated attachment for 3-4 passages; there were 75%-80% 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.