Lejeune H, Skalli M, Sanchez P, Avallet O, Saez J M
INSERM U.307, Hôpital Debrousse, Lyon, France.
Int J Androl. 1993 Feb;16(1):27-34. doi: 10.1111/j.1365-2605.1993.tb01149.x.
An in-vitro method was developed to study Sertoli-Leydig cell interactions in man, using testes removed at the time kidneys were removed for transplantation from 6 young adult men (aged 17-45 years) after cerebral death. After collagenase digestion of testicular tissue, Leydig cells were purified on discontinuous Percoll gradients. Two fractions of Leydig cells, 'L2' and 'L3' which differed in their buoyant density (1.05 g < L2 < 1.06 g and 1.06 g < L3 < 1.08 g), were obtained. The Sertoli cell-enriched preparation was obtained from seminiferous tubular fragments after sequential treatment with glycine buffer to remove peritubular-myoid cells, a second collagenase digestion, mechanical fragmentation and washes to remove germ cells. Purified Leydig cells were then cultured either alone or together with Sertoli cells in culture dishes coated with collagen, fibronectin and laminin in a chemically defined medium without serum. The influence of co-culture on basal testosterone secretion was examined in 3 successive 48 h periods.(ABSTRACT TRUNCATED AT 250 WORDS)
开发了一种体外方法来研究人类睾丸支持细胞-间质细胞的相互作用，使用的是从6名年轻成年男性（年龄17 - 45岁）脑死亡后因肾脏移植而切除的睾丸。对睾丸组织进行胶原酶消化后，在不连续的 Percoll 梯度上纯化间质细胞。获得了两部分间质细胞，“L2”和“L3”，它们的浮力密度不同（1.05 g < L2 < 1.06 g 且 1.06 g < L3 < 1.08 g）。通过用甘氨酸缓冲液依次处理以去除睾丸肌样细胞、第二次胶原酶消化、机械破碎和洗涤以去除生殖细胞，从生精小管片段中获得富含支持细胞的制剂。然后将纯化的间质细胞单独或与支持细胞一起培养在涂有胶原蛋白、纤连蛋白和层粘连蛋白的培养皿中，置于无血清的化学成分确定的培养基中。在连续3个48小时的时间段内检测共培养对基础睾酮分泌的影响。（摘要截断于250字）
