Shi Min, Li Xian-xin, Song Bo, Long Yun, Gui Yao-ting, Cai Zhi-ming
Laboratory of Male Reproduction, Center of Reproductive Medicine, Peking University Shenzhen Hospital, Shenzhen 518036, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2006 Aug 18;38(4):441-3.
To establish the method to identify human testicular spermatogenic cells.
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.
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.
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.
建立鉴定人睾丸生精细胞的方法。
对梗阻性无精子症患者的睾丸活检样本进行机械解离以分散细胞。对混合细胞涂片进行Diff-Quik染色。在配备霍夫曼调制对比光学系统的倒置显微镜下观察活细胞，并根据其形态进行分类。对分类后的细胞涂片应用使用17号染色体着丝粒探针的显色原位杂交（CISH）和抗c-kit免疫细胞化学染色。
在霍夫曼光学系统下可分类的主要四类圆形细胞群为支持细胞、初级粗线期精母细胞（PPS）、精原细胞、圆形精子细胞。在CISH中，支持细胞、PPS和精原细胞显示2个着丝粒信号，而圆形精子细胞和长形精子细胞/精子显示1个着丝粒信号。在免疫细胞化学染色中，PPS和精原细胞显示阳性染色，而支持细胞、圆形精子细胞和精子显示阴性染色。
分散的人睾丸细胞在活细胞/染色形态、倍性分析、细胞表面膜抗原表达方面表现出不同特征。所有这些方法均可用于鉴定不同阶段的人睾丸生精细胞，而在霍夫曼光学系统下进行细胞形态分类是一种简单有效的活细胞鉴定方法。
