Isolation of sertoli cells from adult rat testes: An approach to ex vivo studies of sertoli cell function

Matthew D. Anway, Janet Folmer, William W Wright, Barry R Zirkin

Research output: Contribution to journalArticle

Abstract

Much of what is known about the molecular regulation and function of adult Sertoli cells has been inferred from in vitro studies of immature Sertoli cells. However, adult and immature cells differ in significant ways and, moreover, many Sertoli cell functions are regulated by conditions that are difficult to replicate in vitro. Our objective was to develop a procedure to isolate Sertoli cells rapidly and in sufficient number and purity to make it possible to assess Sertoli cell function immediately after the isolation of the cells. The isolation procedure described here-in takes less than 4 h and does not require culturing the cells. From a single 4-mo-old adult rat, we routinely obtain 7.0 ± 0.4 × 106 Sertoli cells per testis, and from a 21-mo-old rat, 7.2 ± 0.4 × 106 Sertoli cells per testis. The purity, determined by morphologic analyses of plastic-embedded cells or after staining for tyrosine-tubulin or vimentin, averaged 80%. The contaminants typically included germ cells (10%) and myoid cells (10%). The germ cell-expressed genes protamine-2 and hemiferrin were not detected in the Sertoli cell preparations by Northern blot analyses, but the Sertoli cell-expressed genes clusterin, cathepsin L, and transferrin were highly expressed. Transferrin mRNA levels were greater in Sertoli cells isolated from aged than from young adult rats, consistent with previous analyses of whole testes; and cathepsin L mRNA levels were far more highly expressed in Sertoli cells isolated from stages VI-VII than from other stages of the cycle of the seminiferous epithelium, also consistent with previous analyses of whole testes and isolated tubules. These studies indicate that the freshly isolated cells retain differentiated function, and thus it should be possible to assess the in vivo function of adult Sertoli cells by isolating the Sertoli cells and immediately assessing their function.

Original languageEnglish (US)
Pages (from-to)996-1002
Number of pages7
JournalBiology of Reproduction
Volume68
Issue number3
DOIs
StatePublished - Mar 1 2003

Fingerprint

Sertoli Cells
Testis
Cathepsin L
Transferrin
Germ Cells
Clusterin
Seminiferous Epithelium
Messenger RNA
Cell Separation
Vimentin
Northern Blotting
Genes
Plastics
Young Adult

Keywords

  • Aging
  • Sertoli cells
  • Spermatogenesis

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Embryology

Cite this

Isolation of sertoli cells from adult rat testes : An approach to ex vivo studies of sertoli cell function. / Anway, Matthew D.; Folmer, Janet; Wright, William W; Zirkin, Barry R.

In: Biology of Reproduction, Vol. 68, No. 3, 01.03.2003, p. 996-1002.

Research output: Contribution to journalArticle

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