中国生物工程杂志

The objective of this study was to develop a practical in vitro culture system of chicken fibroblasts to facilitate further studies on the feeder cells of chicken embryonic stem cells. Explant techniques and monolayer culture in culturing chicken fibroblasts were compared, the primary chichen fibroblasts derived from tissue explant culture grown slowly and confluented together by 6～7 days of culture, while cells derived from enzymatic digestion grown faster and confluented together by 5 days of culture. However the growing speed of passaged fibrlblasts derived from the two methods was similar, which could confluent within 2 to 3 days of culture. Chicken embryonic tissue digested by 0.25% trypsin provided a high yield of viable proliferating for primary culture. 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 three or four subcultures. There were 75%-80% of cells survived after frozen and thawed. The growth curves of fetal and skin fibroblasts were normal and similar. Chicken embryonic and skin fibroblasts could subculture 12th generation and still retained the normal chromosome content.