[Isolation, cultivation, identification and functional study of fetal mice Leydig cells in vitro].
OBJECTIVE
To explore the methods of isolation, cultivation, purification, identification of the fetal mice testis Leydig cell and to observe its biological characteristics in vitro.


METHODS
Leydig cells were isolated by 0.03% collagenase (type I) from fetal mice testis and cultured in DMEM/F12 medium. The identity and purity of Leydig cell were assessed by 3beta-hydroxysteroid dehydrogenase delta4-delta5 isomerase (3beta-HSD). Cell viability was measured by trypan blue. Testosterone level in the medium of cultured Leydig cells was measured in various culture phases and cell density by radioimmunoassay.


RESULTS
The purity of Leydig cell was (45.10 +/- 1.66)% before culture, and (81.17 +/- 2. 32)% 72 h after culture. The level of testosterone secreted by Leydig cells could be detected in the medium and its level was associated with the density and time of cultured Leydig cells. The secretion capacity of testosterone by single Leydig cell decreased gradually during the culturing period.


CONCLUSIONS
The fetal Leydig cells isolated from fetal mice testis have high purity. It can be cultured and kept the secretion ability of testosterone for a few days in vitro. This system can provide a valuable model for further study on the cellular function of the Leydig cells of fetal mice.
Department of Urology, Children's Hospital, Chongqing University of Medical Science, Chongqing 400014, China.
Xiao-feng
XF
Song
Guang-hui
GH
Wei
Yong-ji
YJ
Deng
D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D007985:Leydig Cells / Q000166:cytology* / Q000502:physiology*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D013737:Testis / Q000196:embryology*; D013739:Testosterone / Q000378:metabolism
D013739:Testosterone