用Percoll分离的大鼠睾丸间质细胞对绒毛膜促性腺激素刺激的反应
The response of leydig cells isolated by Percoll to the stimulation of human Chorionic Gonadotropin
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摘要: 目的:了解用分离液Percoll分离的Leydig细胞对人绒毛膜促性腺激素刺激的反应规律。方法:将大鼠睾丸间质细胞混悬液用不连续梯度(20%～80%)Percoll分离液分离并进行为期5d的培养,观察不同培养时间Leydig对不同剂量hCG刺激的反应。结果:Percoll分离、纯化的Leydig细胞纯度可达80%～85%;随着培养时间的延长,Leydig细胞功能逐渐减低,对hCG刺激反应以培养d1和d2最大;小剂量hCG(1 IU/ml)即可使睾酮分泌达到峰值,加大剂量则会抑制细胞分泌睾酮的功能。结论:Percoll分离液能明显的提高Leydig细胞纯度;培养的d1～2为最佳刺激时间,hCG用量以1 IU/ml为宜Abstract: Objective: To observe the response of Leydig cells isolated by Percoll to the stimulation of human Chorionic Gonadotropin. Methods: Leydig cells were isolated and purified by a discontinuous Percoll gradient of 20%~80% and then cultured for 5 days. human Choronic Gonadotropin was given in different time and in different dosage. Testosterone was measured. Results: The purity of Leydig cells was 80%~85%. Testosterone decreased with time going. The peak of testosterone stimulated by hCG was in the first and second day of culturing. Low dose of hCG could cause highest production of testosterone. High dose of hCG may inhibit secreting of Testosterone. Conclusions:Highly purified Leydig cells could be obtained with Percoll. Both the first and second day of cluturing were the best time to stimulate with hCG. The available dosage of hCG was 1 IU/ml.
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