Establishment of Primary Culture of Sertoli Cells

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


The successful isolation and culture of Sertoli cells depend on a series of delicate processes of mechanical isolation and enzymatic digestion of the testicular tissue, taking advantage of an array of enzymes (such as DNAse, collagenase, and pancreatin) in order to digest the extracellular matrix components. The complexity of these processes may present some differences depending on the origin of the testicular sample (whole tissue or biopsy) and of the species in question. Rat and mouse Sertoli cells are obtained by a similar protocol, whereas bovine and human Sertoli cells require a more extensive mechanical and enzymatic processing.


Enzymatic digestion Mechanical dispersion Primary cell culture Sertoli cells 



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