摘要
用DME:Ham's F12(1∶1)培养液,添加3个水平的表皮生长因子和2个水平的胰岛素,组合成6种 培养体系(CS)分别培养大熊猫皮肤成纤维细胞。通过对细胞生长速度和染色体数目变异率进行测定,测得在 添加10μg／mL的胰岛素和40 ng／mL的表皮生长因子的培养体系(CS-5)中:以(1．673±0．185)×105／mL密 度接种细胞,经3．5 d,密度达到6．890×105／mL,其生长速度最快;染色体数目为二倍体细胞的比率75．77％; 核型分析显示,培养的细胞是大熊猫体细胞。综合衡量,CS-5更适合大熊猫皮肤成纤维细胞的培养。
Six culturing systems were designed by adding different concentrations of epidermal growth factor （EGF） and insulin into DME/Ham＇s F12 medium. Based on the speed of cell growth and variation ratio of chromosome number, we chose the best culture media from the six culture systems. The results showed that the culture medium of 10μg/mL insulin and 40 ng/mL EGF was most suitable for giant panda＇s cutaneous fibroblast culture in vitro. In the best culturing system, the density of fibroblast increased from （ 1. 673 ± 0.185） × 10^5/mL to 6. 890 × 10^5/mL for three and a half days. The fibroblast growing speed was the fastest in all culture system and the ratio of diploid at the 5th generation was 75.77%. Karyotype analysis showed that culture cell is from Giant Panda＇s body cell. In conclusion, CS-5 is more suitable for giant panda＇s cutaneous fibroblast culture than other culturing systems in vitro.
基金
成都大熊猫繁育研究基金(2001003)
