[Identification of human testicular spermatogenic cells at different stages].
OBJECTIVE
To establish the method to identify human testicular spermatogenic cells.


METHODS
Cells were dispersed by mechanic disintegration on testicular biopsy samples of obstructive azoospermic patients. Diff-Quik staining was applied to mixed cell smears. Live cells were observed under inverted microscope equipped with Hoffman modulation contrast optics, and classified according to their morphology. Chromogenic in situ hybridization (CISH) using chromosome 17 centromere probe and anti-c-kit immunocytochemistry staining were applied to classified cell smears.


RESULTS
Sertoli cell, primary pachytene spermatocyte (PPS), spermatogonia, round spermatid were the main four round cell groups that can be classified under Hoffman optics. In CISH, Sertoli cell, PPS and spermatogonia displayed 2 centromere signals, while round spermatid and elongated spermatid/sperm displayed 1 centromere signal. In immunocytochemistry staining, PPS and spermatogonia displayed positive staining, while Sertoli cell, round spermatid and sperm displayed negative staining.


CONCLUSIONS
Dispersed human testicular cells displayed different characteristics in live/staining morphology, ploidy analysis, cell surface membrane antigen expression. All of these methods can be chosen to identify human testicular spermatogenic cells at different stages, while cell morphology classifying under Hoffman optics is a simple and effective method for live cell identification.
Shi
Min
M
Laboratory of Male Reproduction, Center of Reproductive Medicine, Peking University Shenzhen Hospital, Shenzhen 518036, China.
Li
Xian-xin
XX
Song
Bo
B
Long
Yun
Y
Gui
Yao-ting
YT
Cai
Zhi-ming
ZM
D000328:Adult; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000379:methods; D008297:Male; D012708:Sertoli Cells / Q000166:cytology / Q000378:metabolism; D013087:Spermatids / Q000166:cytology* / Q000378:metabolism; D013090:Spermatocytes / Q000166:cytology / Q000378:metabolism; D013091:Spermatogenesis; D013093:Spermatogonia / Q000166:cytology / Q000378:metabolism; D013094:Spermatozoa / Q000166:cytology* / Q000378:metabolism; D013737:Testis / Q000166:cytology* / Q000378:metabolism