Viability of meiotic prophase spermatocytes of rats is facilitated in primary culture of dispersed testicular cells on collagen gel by supplementing epinephrine or norepinephrine: Evidence that meiotic prophase spermatocytes complete meiotic divisions in vitro
- 1 December 1989
- journal article
- research article
- Published by Springer Nature in In Vitro Cellular & Developmental Biology
- Vol. 25 (12) , 1088-1098
- https://doi.org/10.1007/bf02621259
Abstract
Dispersed testicular cells prepared from 14-d-old rats were cultured on type 1 collagen gels using a medium composed of a 1∶1 mixture of Ham’s F12 medium and Leibovitz’s L15 medium (F12-L15 medium) containing 10% (vol/vol) fetal bovine serum. The viability of the spermatogenic cells was facilitated by supplementing a rat adrenal extract into the medium. The effective substance(s) (the survival factor) was purified from acid extracts of adrenals by molecular sieve high performance liquid chromatography and identified as epinephrine and norepinephrine. Both epinephrine and norepinephrine promoted the survival of the spermatogenic cells with a half saturating dose of 10 ng/ml. The spermatogenic cells, which could be cultured for 2 wk on a collagen gel by supplementing with the survival factor (epinephrine or norepinephrine), were subjected to Giemsa staining and to DNA flow cytometry. The following results were obtained: a) The spermatogenic cells from 14-d-old rats did not contain spermiogenic cells (lc-cells). b) During a culture period of 2 to 7 d the ratio of meiotic prophase spermatocytes (4c-cells) to premeiotic cells (2c-cells) increased. On Day 7, more than 90% of the surviving cells were meiotic prophase spermatocytes. c) On Day 10, spermatids (lc-cells) appeared for the first time. The time of the first appearance of spermatids in the culture was consistent with that in vivo. These results suggest that both epinephrine and norepinephrine facilitated the viability of meiotic prophase spermatocytes and that a part of the meiotic prophase spermatocytes completed the meiotic divisions in the testicular cell culture.This publication has 33 references indexed in Scilit:
- β-Adrenergic receptors and stimulatory effects of (−) isoproterenol on testosterone production in fetal mouse Leydig cellsBiochemical and Biophysical Research Communications, 1988
- Influence of adrenalectomy on maturation of gonadotrophin function in the male ratJournal of Endocrinology, 1982
- Catecholamine stimulation of testosterone production via cyclic AMP in mouse leydig cells in monolayer cultureMolecular and Cellular Endocrinology, 1982
- Identification and characterization of a β1-adrenergic receptor in the rat sertoli cellMolecular and Cellular Endocrinology, 1981
- Secretion of plasminogen activator by Sertoli cell enriched culturesMolecular and Cellular Endocrinology, 1977
- Spermatogenic cells of the prepuberal mouse: isolation and morphological characterizationThe Journal of cell biology, 1977
- Separation of mouse spermatogenic cells by sedimentation velocityDevelopmental Biology, 1976
- Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining.The Journal of cell biology, 1975
- Stimulatory effect of vitamins and glutamine on the differentiation of germ cells in rat testes organ culture grown in chemically defined mediaExperimental Cell Research, 1966
- In vitro culture of rat testicular cellsExperimental Cell Research, 1966