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Evaluation of the Purity of Sertoli Cell Primary Cultures

  • Raquel L. Bernardino
  • Marco G. Alves
  • Pedro F. OliveiraEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1748)

Abstract

The purity of the primary cultures of Sertoli cells is a factor for the validation of the studies using this methodological approach. There is a probability of contamination of these cultures with other testicular cellular types, such as peritubular cells and germ cells (that represent a large percentage of the volume of the seminiferous tubules).

For the evaluation of the purity of cultures, the immunocytochemistry technique (immunoperoxidase or immunofluorescence) is frequently used to label a protein marker specific for the Sertoli cells within the testicular environment. The expression of several protein markers can be used, with different particularities, being that vimentin is often used as a marker for Sertoli cells. Vimentin is expressed independently of the differentiation state of the cells and is observed around the nuclei and in the cytoplasm of Sertoli cells.

Keywords

Antibody Immunocytochemistry Immunofluorescence Immunoperoxidase Prepubertal stage Culture purity Sertoli cells Vimentin 

Notes

Acknowledgments

This work was supported by the “Fundação para a Ciência e a Tecnologia” (FCT) (PTDC/BBB-BQB/1368/2014) and UMIB (PEst-OE/SAU/UI0215/2014). R.L. Bernardino was also financed by FCT (SFRH/BD/103105/2014).

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Raquel L. Bernardino
    • 1
  • Marco G. Alves
    • 1
  • Pedro F. Oliveira
    • 2
    • 3
    • 4
    • 5
    Email author
  1. 1.Unit for Multidisciplinary Research in Biomedicine (UMIB), Laboratory of Cell Biology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar (ICBAS)University of PortoPortoPortugal
  2. 2.Department of Microscopy, Laboratory of Cell Biology and Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute of Biomedical Sciences Abel Salazar (ICBAS)University of PortoPortoPortugal
  3. 3.Department of Genetics, Faculty of MedicineUniversity of PortoPortoPortugal
  4. 4.i3S - Instituto de Investigação e Inovação em SaúdeUniversity of PortoPortoPortugal
  5. 5.Department of Biosciences, Biotechnologies and BiopharmaceuticsUniversity of Bari “Aldo Moro”BariItaly

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