摘要
本文介绍了用胰蛋白酶分散大鼠黄体细胞和培养的方法。未成年雌性大鼠用PMSG和hCG诱导超排卵后取得的黄体,经0.025%的胰蛋白酶分散,可获得大(直径为14.2～28.5μm)、小(直径为7.1～12.3μm)二种黄体细胞。台盼兰排斥试验测得细胞活力为85～90%,经3～4小时孵育后基本保持不变。黄体细胞产生的孕酮量与hCG浓度、培养时间和细胞数明显相关;而且,cAMP能显著增加hCG刺激的孕酮产生。结果表明,用胰蛋白酶分散大鼠黄体细胞进行体外培养是一种简单而有用的方法。
A method for the isolation and incubation of rat luteal cells in vitro is introduced. Corpora lutea were obtained from immature rat pretreated with PMSG and hCG 7 days after hCG injection. Corpora lutea were dissociated by 0.025% trypsin and the isolated luteal cells were washed three times with Kreb-Ringer bicarbonate glucose buffer containing 0.5% bovine serum albumin (KRBGAL). The suspension contained luteal cells of two sizes; the diameter of large cells was 14.2～28.5 μm and small cells 7.1-12.3 μm. Viability of the recovered cells was generally 85～90% as judged by Trypan blue exclusion test and did not change markedly during four hours incubation. The luteal cells could produce progesterone, and the progesterone production was related to concentration of hCG added to the media, the time of incubation and the number of cells, cAMP can increase hCG-stimulated progesterone production singnificantly. When the number of cells was given, after incubation for three to four hours, the basal and hCG-stimulated progesterone production was constant. These results show that trypsin digestion to isolate rat luteal cells for in vitro study is a simple and useful method.
出处
《生殖与避孕》
CAS
CSCD
北大核心
1989年第2期57-60,15,共4页
Reproduction and Contraception
