Davebrid: updated with more details

2012-09-05T15:50:26Z

updated with more details

@@ Line 1: / Line 1: @@
 From Zolov et al (under review)
 ==Generation of Mouse Primary Fibroblast Cells==
-Fibroblasts were prepared from tails and legs of P0 pups. Tissues from individual animals were kept separate, disaggregated with a surgical blade and digested with 500 U/ml type II collagenase (LS004204, Worthington, US) in 5 ml RPMI media (Cat. # 1640, Invitrogen, US) supplemented with 10% FBS, 2 mM L-glutamine and penicillin/streptomycin for 12 hours at 37°C. After incubation, cells were harvested at 200 g for 5 min and plated in 100 mm dishes, in fresh RPMI medium supplemented with 10% FBS, 2 mM L-glutamine and penicillin/streptomycin. Experiments were performed on cells that were passaged 1-3 times.
+# MEFs were prepared from tails and legs of P0 pups.

Davebrid: copied in protocol from sergey's paper

2012-09-04T16:32:42Z

copied in protocol from sergey's paper

New page

From Zolov et al (under review)

==Generation of Mouse Primary Fibroblast Cells==

Fibroblasts were prepared from tails and legs of P0 pups. Tissues from individual animals were kept separate, disaggregated with a surgical blade and digested with 500 U/ml type II collagenase (LS004204, Worthington, US) in 5 ml RPMI media (Cat. # 1640, Invitrogen, US) supplemented with 10% FBS, 2 mM L-glutamine and penicillin/streptomycin for 12 hours at 37°C. After incubation, cells were harvested at 200 g for 5 min and plated in 100 mm dishes, in fresh RPMI medium supplemented with 10% FBS, 2 mM L-glutamine and penicillin/streptomycin. Experiments were performed on cells that were passaged 1-3 times.

Isolation of Mouse Embryonic Fibroblasts (MEFs) - Revision history

Davebrid: updated with more details

Davebrid: copied in protocol from sergey's paper