title: A new protocol for cultivation of predegenerated adult rat Schwann cells.
abstract: The purpose of this study was to optimize the methodology of cultivation of predegenerated Schwann cells (SCs). SCs were isolated from 7-day-predegenerated sciatic nerves of adult rats. We applied commercially available culture medium for cultivation of endothelial cells endothelial cell culture medium (EBM-2) instead of Dulbecco's Modified Eagle's Medium commonly used to culture adult Schwann cells. Additionally, cell culture medium was supplemented with factors specifically supporting SCs growth as: bovine pituitary extract (5 μg/ml), heregulin (40 ng/ml) and insulin (2.5 ng/ml). Similarly to the reports of others authors, we did not observe any beneficial effects of Forskolin application, so we didn't supplement our medium with it. Cell culture purity was determined by counting the ratio of GFAP, N-Cadherin and NGFR p75-positive cells to total number of cells. About 94-97 % of cells were confirmed as Schwann cells. As a result, we obtained sufficient number and purity of Schwann cells to be applied in different experimental models in rats. EBM-2 medium coated with fibronectin was the best for cultivation of adult rat Schwann cells.
authors.affiliation: Department of Physiology, Medical University of Silesia, ul. Medyków 18, 40-752, Katowice, Poland, maritadutczak@gazeta.pl.
authors.forename: Marita
authors.initials: M
authors.lastname: Pietrucha-Dutczakv
authors.forename: Wiesław
authors.initials: W
authors.lastname: Marcol
authors.forename: Tomasz
authors.initials: T
authors.lastname: Francuz
authors.forename: Dariusz
authors.initials: D
authors.lastname: Gołka
authors.forename: Joanna
authors.initials: J
authors.lastname: Lewin-Kowalik
mesh_terms: D000375:Aging; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured / Q000378:metabolism; D003470:Culture Media; D051381:Rats; D012583:Schwann Cells / Q000166:cytology*; D012584:Sciatic Nerve / Q000166:cytology*
chemical_list: D003470:Culture Media