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A New Protocol for Cultivation of Predegenerated Adult Rat Schwann Cells

A New Protocol for Cultivation of Predegenerated Adult Rat Schwann Cells

Marita Pietrucha-Dutczakv, Wiesław Marcol, Tomasz Francuz, Dariusz Gołka, Joanna Lewin-Kowalik

Cell Tissue Bank. 2014 Sep;15(3):403-11.

PMID: 24197905

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42413247	Endothelial cell growth supplement from bovine pituitary			Price

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