Fibronectin synthesis is a marker for peritubular cell contaminants in Sertoli cell-enriched cultures
With indirect immunofluorescent microscopic techniques, we have shown that fibronectin is distributed primarily in or along the basal lamina of the seminiferous tubule boundary tissue in sections of testes from 20-day-old rats. Purified rat Sertoli cell-enriched aggregates, maintained in culture in...
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Published in | Biology of reproduction Vol. 30; no. 1; pp. 199 - 211 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
Madison, WI
Society for the Study of Reproduction
01.02.1984
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Subjects | |
Online Access | Get full text |
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Summary: | With indirect immunofluorescent microscopic techniques, we have shown that fibronectin is distributed primarily in or along
the basal lamina of the seminiferous tubule boundary tissue in sections of testes from 20-day-old rats. Purified rat Sertoli
cell-enriched aggregates, maintained in culture in the presence or absence of serum, exhibit no detectable immunofluorescence
with fibronectin antibody, whereas purified peritubular cells in culture do have a positive reaction to fibronectin antibody.
Peritubular cells in culture incorporate [35S] methionine into fibronectin which can be immunoprecipitated with a fibronectin
antiserum, but Sertoli cells do not. We have used various criteria to estimate the degree of purity of Sertoli cell-enriched
preparations. The presence of peritubular myoid cells in conventional Sertoli cell-enriched aggregates, cultured in the presence
or absence of serum, can be detected with transmission electron microscopic examination, by the Feulgen staining procedure,
and by the immunocytochemical identification of fibronectin. We describe a technique to purify Sertoli cells in conventional
Sertoli cell-enriched preparations by treatment with hyaluronidase, resulting in a lesser number of peritubular cells by the
above criteria, even in preparations cultured in the presence of serum. Data presented suggest that some of the products previously
attributed exclusively to Sertoli cells in Sertoli cell-enriched preparations, particularly those cultured in the presence
of serum, may have been contributed by peritubular cells. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0006-3363 1529-7268 |
DOI: | 10.1095/biolreprod30.1.199 |