Etchevers:Notebook/Embryos/2009/06/15

From OpenWetWare
Jump to: navigation, search
C14.jpg Recueil d'embryons Purpan <html><img src="/images/9/94/Report.png" border="0" /></html> Recueil d'embryons Purpan - principal
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Reseed DRG cells on collagen-coated coverslips

After discussion with Michele Allouche Friday, decided to compare the effects of her neural differentiation medium on the DRG cultures.

Seeded 5K, 10K and 15K of each of EMB050003 and EMB050005 DRG cells (passage 4) on 1.5h collagen-coated, 70% EtOH-sterilized glass coverslips, rinsed with PBS. 4 coverslips each, to be 2 each in RICH or neural media as of tomorrow. (Cells on 2 cm2.)

Fresh Rich media does not have any FGF2 in it. For the overnight in the 24-well plate, 1/4 old Rich medium, which did.

EMB03 had 1,220,000 cells and EMB05 had 1,630,000 cells.

Diluted into 4 mL (1 mL per well): 3, 6 and 9 μL or 4, 8 and 12 μL respectively, to get appropriate plating densities.

Fixed the highly confluent 8-well slides that were seeded with each of these cultures last Thursday. 30 minutes RT PAF, rinsed in PBS +Ca/Mg2, now in refrigerator.

Re-plated the residual DRG cells in 75 cm2 flasks, no collagen, to see if they adhere on their own.