The whole meiotic process can occur in vitro in untransformed rat spermatogenic cells
| Autor: | M.H. Perrard-Sapori, Dominique Segretain, Dominique Hue, Jean-Claude Nicolle, Christophe Staub, Philippe Durand |
|---|---|
| Přispěvatelé: | ProdInra, Migration, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Communications Cellulaires et Différenciation (CCD), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique (INRA) |
| Jazyk: | angličtina |
| Rok vydání: | 2000 |
| Předmět: |
Male
Chromosomal Proteins Non-Histone [SDV]Life Sciences [q-bio] Cell Population Spermatocyte Biology [INFO] Computer Science [cs] Andrology 03 medical and health sciences 0302 clinical medicine Meiosis Culture Techniques medicine Animals [INFO]Computer Science [cs] RNA Messenger Rats Wistar Spermatogenesis education ComputingMilieux_MISCELLANEOUS DNA Primers 030304 developmental biology 0303 health sciences education.field_of_study 030219 obstetrics & reproductive medicine Base Sequence Spermatid Cell Biology Spermatozoa Molecular biology In vitro Rats [SDV] Life Sciences [q-bio] Microscopy Electron Chemically defined medium medicine.anatomical_structure Bromodeoxyuridine CULTURE DE CELLULE RAT |
| Zdroj: | Experimental Cell Research Experimental Cell Research, Elsevier, 2000, 260, pp.85-95 |
| ISSN: | 0014-4827 1090-2422 |
| Popis: | The aim of the present study was to assess whether the whole meiotic process of spermatogenic cells is able to take place in vitro. Fragments of seminiferous tubules from 20- to 22- or 28-day-old rats were seeded in medium containing 0.2% fetal calf serum in bicameral chambers and then cultured for 4 weeks in a chemically defined medium. The differentiation of meiotic germinal cells was followed by four criteria: (i) ultramicroscopic examination of the different types of germ cells present in the cell layer throughout the culture period; (ii) determination of the changes in DNA content per nucleus of the cell population seeded with time in culture; (iii) assessment of the ability of germinal cells to transcribe genes expressed after completion of meiosis; and (iv) monitoring the fate of BrdU-labeled leptotene spermatocytes. The ultrastructural study showed that the overall organization of the cells in the culture well recalls that of the seminiferous epithelium throughout the culture period. Moreover the identification of young round spermatids 21 days after seeding suggested that these spermatids had been formed very recently in culture. Determination of DNA content per nucleus showed that a 1C cell population could be observed after several days of cultures reaching 6 to 10% of total cells. An exponential-like increase in the amounts of the mRNAs encoding for TP1 or TP2 occurred from the time when 1C cells appeared in the culture until the end of the experiment. Finally, BrdU-labeled leptotene spermatocytes differentiated into pachytene spermatocytes and then into secondary spermatocytes, and BdrU-labeled round spermatids were observed from Day 21 of culture onward. Taken together these results indicate that the whole meiotic process from leptotene spermatocyte to round spermatid can indeed occur in vitro under the present culture conditions. |
| Databáze: | OpenAIRE |
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