埃及伊蚊中肠上皮细胞的原代培养和细胞的持续培养(英文)

ESTABLISHMENT OF PRIMARY AND CONTINUOUS CULTURE OF MIDGUT EPITHELIAL CELLS FROM AEDES AEGYPTI (DIPTERA:CULICIDAE) IN VITRO

目的 探讨蚊虫中肠上皮细胞的培养 ,建立细胞的原代培养和细胞持续培养。 方法 用含抗生素的 10 %蔗糖水喂养新羽化出的埃及伊蚊成蚊 ,3d后 ,在无菌的条件下解剖中肠 ,用含抗生素的平衡盐溶液洗 3次 ,放入Grace’s培养基中 (含有 7%小牛血清 ,0 .5 %庆大霉素 ,1mg/ml抗酵母菌素和 6μl/ml复合维生素 ) ,2 5℃培养箱中培养。 结果 培养 3d后 ,95 %的细胞生长良好 ,7d后 ,大约 2 5 %的细胞死亡。第 3d的细胞死亡率明显低于以后的任何时间 (P <0 .0 5 ) ,从第 7d到 3 5d细胞死亡率差异无显著性 (P >0 .0 5 )。成功地建立起中肠上皮细胞的原代培养并能持续存活 40多天 ,保持细胞的连续培养。 结论 所发展的埃及伊蚊中肠上皮细胞的原代培养和细胞的持续培养的方法是成功的。

Objective To investigate the growth and development of the adult mosquito midgut, to develop primary midgut culture. Methods After emerging adult mosquitoes were fed for 3 consecutive days on antibiotic solution, midguts dissected under sterile condition were kept in Grace's medium containing 7% fetal bovine serum, 0.5% gentamycin, 1 mg/ml yeastolate and 6 μl/ml vitamin premix to incubate at 25℃. Results The cells recovered from the midgut are healthy with about 95% viability on day 3. Approximately 25% of the cells died one week later, but those that survive stay in a stable condition during 35 days. There are significantly fewer dead cells on day 3 ( P < 0.05 ) than at any time during the remaining 32 days of incubation. There is no statistical difference in cell mortality from day 7 to day 35 ( P > 0.05 ). Primary cultures that would survive up to over a 40-day incubation period were successfully developed and continuous cultures were established. Conclusion The procedure that would facilitate the culture of midgut epithelial cells from Aedes aegypti adult mosquitoes was developed.