Preparation of spermatogenic cell populations at specific stages of differentiation in the human

Molecular Reproduction and Development - Tập 30 Số 3 - Trang 275-282 - 1991
Yannick Blanchard1, Marie-Thérèse Lavault2, D. Quernée2, D. Le Lannou3, Bernard Lobel4, D. Lescoat2
1CECOS de l’Ouest, Rennes, France
2Laboratoire d'Histologie et d'Embryologie, France
3CECOS de l'Ouest, France
4Service d'Urologie, Rennes, France

Tóm tắt

AbstractStudying biochemical events in human spermatogenesis requires separated populations of spermatogenic cells. Dissociation of these cells was performed by a Trypsin‐DNAse method adapted from the technique used for rodents. Cell separation was performed by centrifugal elutriation. Seven populations were collected, one further purified by Percoll gradient centrifugation, giving nine different cell populations. The efficiency of the cell separation was evaluated by phase contrast microscopy, flow cytometric DNA analysis, and electron microscopy. Five populations were enriched in spermatids: two in round spermatids (87% and 73%), another in round (52%) and elongating (44%) spermatids, another constituted by 80% elongating spermatids, and the last by 90% elongated spermatids. Two of the four remaining populations were enrichied in primary spermatocytes (74% and 54%); another population was the upper part of the Percoll gradient and constituted cytoplasmic lobes and residual bodies (89%); the last population was made up of various cells, with no specific enrichment. Electron microscopic observations revealed good preservation of the separated cells; only the flagella from elongated spermatids were lost. Furthermore, an unusual pattern of nucleoplasm distribution during stages 2–4 of spermatid differentiation was observed and its signification is discussed with regard to the shape of the human spermatozoon.

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Molecular Reproduction and Development

Tập 30 Số 3

275-282

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