水牛皮肤成纤维细胞的分离与体外培养

In Vitro Isolation and Culture of Buffalo (Bubalus bubalis) Skin Fibroblast Cells

探讨水牛成纤维细胞的分离与传代培养方法。组织块培养法培养的成纤维细胞原代生长较慢,需12天左右方可汇合形成单层,而酶消化法培养的成纤维细胞原代生长相对生长快,仅需8天便可汇合形成单层。两种方法传代细胞的生长速度相似,仅需4￣5天就可汇合形成单层。通过体细胞的核型分析发现,成纤维细胞在传代培养过程中的核型变化不大,66.67%￣81.67%的细胞具有正常的二倍体核型,各代之间无显著差异。结果表明,水牛成纤维细胞均能稳定地进行传代培养。

Methods for isolation and culture of buffalo fibroblast cells were studied. The primary buffalo fibroblasts derived from tissue explant culture grown slowly and confluented together by 12 days of culture, while fibroblasts derived from enzymatic digestion grown faster and confluented together by 8 days of culture. However, the growing speed of passaged fibroblasts derived from the two methods was similar, which could confluent within 4 to 5 days of culture. There were 70%-80% of cells survived after frozen and thawed. Karyotype analysis showed that 66.67%-81.67% of fibroblasts had normal karyotype, which did not show significant difference among passages. These results indicated that fibroblast cells can be passaged and keep relatively stable karyotype in vitro.