Modified protocol for culturing predegenerated adult rat Schwann cells

• Autorzy: Lewin-Kowalik J. , Marcol W. , Francuz T. , Pietricha-Dutczak M.
• Języki publikacji: EN

Abstrakty

EN

The purpose of this experiment was to optimize the methodology of culturing predegenerated Schwann cells. Right sciatic nerves of adult rats (n=3) were cut and left for 7 days. Then, 1-mm fragments of predegenerated (P) and intact (C) nerves were separately planted in 12-well culture plates precoated with laminin or fibronectin. Medium for culturing of endothelial cells EBM-2 (endothelial cell culture medium) was compared with DMEM (Dulbecco’s Modified Eagle’s Medium). Additionally, culture media were supplemented with factors supporting SCs growth: bovine pituitary extract (5 µg/ml), heregulin (40 ng/ml), and insulin (2.5 ng/ml). After 7 or 14 days, plates were subjected to analysis. Cell culture purity was determined under the fluorescent microscope by estimating the percentage of GFAP, N-Cadherin and NGFR p75-positive cells, and intensity of cell growth - by counting the number of cell islets migrating from nerve explants. Percentage of cells confirmed as Schwann cells was 94–97. Number of islets was significantly higher in both time-frames: (1) in plates precoated with fibronectin in both groups; (2) in P than in C groups. Thus, nerve predegeneration, application of EBM-2 as culture medium and fibronectin as coating appeared a good method for obtaining cultured Schwann cells to be used in different experimental models in rats.

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2013
• Tom: 73
• Numer: Suppl.1
• Opis fizyczny: p.63

Twórcy

autor

Lewin-Kowalik J.

• Department of Physiology,Medical University of Silesia, Katowice, Poland

autor

Marcol W.

• Department of Physiology, Medical University of Silesia, Katowice, Poland

autor

Francuz T.

• Department of Biochemistry, Medical University of Silesia, Katowice, Poland

autor

Pietricha-Dutczak M.

• Department of Physiology, Medical University of Silesia, Katowice, Poland