Woodruff T K, Borree J, Attie K M, Cox E T, Rice G C, Mather J P
Department of Cell Culture Research, Genentech, Inc., South San Francisco, California 94080.
Endocrinology. 1992 Feb;130(2):871-81. doi: 10.1210/endo.130.2.1310280.
Flow cytometry was used to separate and identify Sertoli and germ cell populations in primary rat testicular cultures derived from animals of different ages on the basis of cell size and DNA and lipid content. Multiparameter fluorescent evaluation of each cell preparation resulted in the assignment of specific staining patterns to Sertoli cells (diploid, high lipid content), spermatogonia (diploid, low lipid content), spermatocytes (large, tetraploid, high lipid content), and round spermatids (haploid, low lipid content). Each field was separately analyzed for inhibin and activin binding. Fluorescein isothiocyanate-conjugated activin bound with greatest intensity to spermatogonia, with little binding to leptotene or zygotene spermatocytes. Fluorescein isothiocyanate-conjugated inhibin bound to all stages of germ cells tested. Cross-competition data indicate that at least two and probably three distinct receptors exist for these peptides.
利用流式细胞术，根据细胞大小、DNA和脂质含量，对来自不同年龄动物的原代大鼠睾丸培养物中的支持细胞和生殖细胞群体进行分离和鉴定。对每个细胞制剂进行多参数荧光评估，结果为支持细胞（二倍体，高脂质含量）、精原细胞（二倍体，低脂质含量）、精母细胞（大，四倍体，高脂质含量）和圆形精子细胞（单倍体，低脂质含量）确定了特定的染色模式。对每个视野分别进行抑制素和激活素结合分析。异硫氰酸荧光素偶联的激活素与精原细胞的结合强度最大，与细线期或偶线期精母细胞的结合很少。异硫氰酸荧光素偶联的抑制素与所有测试的生殖细胞阶段都有结合。交叉竞争数据表明，这些肽至少存在两种、可能三种不同的受体。
