Gale J S, Wakefield J S, Ford H C
J Endocrinol. 1982 Feb;92(2):293-302. doi: 10.1677/joe.0.0920293.
A rapid method for preparing Leydig cells from rat testes is described. An interstitial cell suspension, prepared by collagenase treatment of decapsulated testes, was centrifugal for 10 min over a cushion of 60% (v/v) Percoll to remove red blood cells, and then centrifuged for 20 min in a 0-60% linear density gradient of Percoll. Seventy-four per cent of the cells present in that fraction of the gradient comprising 35-50% Percoll were Leydig cells; the yield from each testis was about 1.5 x 10(6) cells. The Leydig cells appeared viable, excluded Trypan blue, possessed high-affinity binding sites for human chorionic gonadotrophin (hCG) and synthesized increased quantities of testosterone in response to hCG. The cells could be stored overnight in 20% (v/v) glycerol at -20 degrees C, with only minimal effect on the specific activities of a number of enzymes used as markers of subcellular components. Testosterone production in vitro by the cells after storage for 20 h was greater than that of hCG-stimulated fresh cells and was not further increased by hCG.
本文描述了一种从大鼠睾丸制备莱迪希细胞的快速方法。通过用胶原酶处理去包膜的睾丸制备间质细胞悬液，在60%（v/v） Percoll垫层上离心10分钟以去除红细胞，然后在0-60%的线性Percoll密度梯度中离心20分钟。在梯度中35-50% Percoll的那一部分中，74%的细胞是莱迪希细胞；每个睾丸的产量约为1.5×10⁶个细胞。莱迪希细胞看起来有活力，排斥台盼蓝，具有人绒毛膜促性腺激素（hCG）的高亲和力结合位点，并在hCG刺激下合成更多的睾酮。细胞可以在-20℃下于20%（v/v）甘油中储存过夜，对用作亚细胞成分标志物的多种酶的比活性只有最小的影响。储存20小时后的细胞体外睾酮产量高于hCG刺激的新鲜细胞，且hCG不再进一步增加其产量。
