Proliferation conditions for human satellite cells. The fractional content of satellite cells

APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica
Michael GasterH D Schrøder

Abstract

Primary satellite cell cultures have become an important tool as a model system for skeletal muscles. A common problem in human satellite cell culturing is fibroblast overgrowth. We combined N-CAM (Leu19) immunocytochemical staining of satellite cells (Sc) with stereological methods to estimate the fraction of Sc in culture. Evaluation of different culture conditions allowed us to find proliferation conditions preferentially for Sc: a) Sc should be cultured on surfaces coated with ECM-gel. b) Primary cell culture should be inoculated in DMEM supplemented with 10% fetal calf serum to increase cell adherence. c) Change of media to DMEM supplemented with 2% Ultroser-G and 2% FCS after 24 h.d) Before subcultivation, cells should be preplated for 30 min. The fractional content of Sc in passage four when applying this method of cultivation was 0.82 +/- 0.07 (mean +/- SE, N = 10). Our method enabled us to establish culture conditions which resulted in high Sc content despite several subcultivations. Estimation of the fractional cell content could be a useful tool for optimizing not only Sc-culturing but all cultures initially containing more cell types.

Citations

Sep 10, 2003·Best Practice & Research. Clinical Endocrinology & Metabolism·Henning Beck-NielsenMichael Gaster
Jun 17, 2009·Diabetes, Obesity & Metabolism·A J McAinch, D Cameron-Smith
Sep 10, 2011·Biochemistry Research International·Nataša NikolićMaria Strömstedt
Dec 29, 2010·Biochemical and Biophysical Research Communications·Michael Gaster
Dec 23, 2009·Biochemical and Biophysical Research Communications·Michael Gaster
Feb 26, 2008·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Ine Blankenberg SkottheimAnders Asberg
Jan 1, 2007·Obesity Research & Clinical Practice·A J McAinchD Cameron-Smith
Jul 30, 2005·Journal of Cellular Biochemistry·Rigmor SolbergJanne E Reseland
Oct 13, 2015·Methods : a Companion to Methods in Enzymology·Mark JuhasNenad Bursac
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Jun 10, 2015·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Craig Robert WrightAaron Paul Russell
Sep 3, 2010·Biochemical and Biophysical Research Communications·Ariane D Minet, Michael Gaster
Dec 3, 2016·The American Journal of Pathology·Louise H JørgensenHenrik D Schrøder
Dec 15, 2018·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Michael Gaster
Jul 17, 2009·American Journal of Physiology. Cell Physiology·Tetsuo YamaguchiYoriko Atomi
Jan 22, 2010·American Journal of Physiology. Cell Physiology·Fred HaugenChristian A Drevon
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Apr 17, 2012·Biochemistry Research International·Michael Gaster
Jul 19, 2017·Journal of Tissue Engineering·Eva Kildall HejbølHenrik Daa Schrøder

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