Progression throughout All Stages of Meiosis from the Early Prophase of Newt Primary Spermatocytes in vitro |
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Authors: | AKIO NISHIKAWA SHIN-ICHI ABE |
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Institution: | Department of Biology, Faculty of Science, Kumamoto University, Kurokami-machi, Kumamoto, 860, Japan |
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Abstract: | Stages of prophase of living primary spermatocytes were determined by use of Rose culture chambers (1). Dissociated primary spermatocytes were cultured at low cell-density in a collagen matrix at 22°C or 27°C and the percentages of cells which had progressed from various stages in prophase through meiosis to various advanced stages were measured. In a standard medium (Leibovitz-15 + 10% fetal bovine serum), more than 70% of the primary spermatocytes at stages beyond the pachytene stage could advance to round spermatids with flagella within a few days at 22°C. The percentages of cells that progressed from stages before the late zygotene stage were less, but at least 13 % of leptotene cells reached metaphase I within a week at 22°C. The percentage of cells that progressed was slightly lower at 27°C than at 22°C: 6.3 and 4.3 days were required for progress from leptotene to metaphase I at 22°C and 27C, respectively. Fetal bovine serum was not indispensable for progression through meiosis. Moreover, 0.5–5.0 μg/ml ovine follicle stimulating hormone (NIAMDD-o-FSH-13), 0.01–1.0 μg/ml 5α-dihydrotestosterone and 1.0 μg/ml testosterone propionate had no significant effect in increasing the percentage of cell progression at 22°C. |
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