Primary Sertoli Cell Cultures From Adult Mice Have Different Properties Compared With Those Derived From 20-Day-Old Animals

Author:

Saewu Arpornrad1,Kongmanas Kessiri12,Raghupathy Riya1,Netherton Jacob3,Kadunganattil Suraj1,Linton James-Jules1,Chaisuriyong Watchadaporn1,Faull Kym F4,Baker Mark A3,Tanphaichitr Nongnuj125ORCID

Affiliation:

1. Chronic Disease Program, Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

2. Department of Biochemistry, Microbiology, Immunology, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, Canada

3. Department of Environmental and Life Science, University of Newcastle, Callaghan, New South Wales, Australia

4. Pasarow Mass Spectrometry Laboratory, University of California, Los Angeles, California

5. Department of Obstetrics and Gynecology, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, Canada

Abstract

Abstract Cultures of Sertoli cells isolated from 20-day-old mice are widely used in research as substitutes for adult Sertoli cell cultures. This practice is based on the fact that Sertoli cells cease to proliferate and become mature in vivo by 16 to 20 days after birth. However, it is important to verify whether cultured Sertoli cells derived from 20-day-old mice do not proliferate ex vivo and whether they have the same properties as cultured adult Sertoli cells. Herein we described an isolation/culture method of Sertoli cells from 10-week-old adult mice with > 90% purity. Properties of these cultured adult Sertoli cells were then compared with those of cultured Sertoli cells derived from 20-day-old mice (also > 90% purity). By cell counting, bromo-2-deoxyuridine incorporation, and metaphase plate detection, we demonstrated that only adult Sertoli cells did not proliferate throughout 12 culture days. In contrast, Sertoli cells derived from 20-day-old mice still proliferated until Day 10 in culture. The morphology and profiles of intracellular lipidomics and spent medium proteomics of the 2 cultures were also different. Cultured adult Sertoli cells were larger in size and contained higher levels of triacylglycerols, cholesteryl esters, and seminolipid, and the proteins in their spent medium were mainly engaged in cellular metabolism. In contrast, proteins involved in cell division, including anti-Mullerian hormone, cell division cycle protein 42 (CDC42), and collagen isoforms, were at higher levels in Sertoli cell cultures derived from 20-day-old mice. Therefore, cultured Sertoli cells derived from 10-week-old mice, rather than those from 20-day-old animals, should be used for studies on properties of adult Sertoli cells.

Funder

Canadian Institutes of Health Research

Development and Promotion of Science and Technology Talented Project

Publisher

The Endocrine Society

Subject

Endocrinology

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