Help: Mouse Dendritic Cells preparation

Mohan mohan_natesan at dfci.harvard.edu
Fri Oct 25 09:52:54 EST 1996


I am trying set up mouse T cell stimulation assays with DCs and
anti-CD3 mAb. I have been unsuccessful in purifying DC. Then main
problem is yield. I am not getting enough cells to complete an assay. I
tried two methods a) Density gradient BSA Path-O-Cyte : I got 2x10 5
cells from 4 spleens b) Metrizamide gradient centrifugation: the yield
was better, but still very low.

1. Can someone suggest a method which will yield respectable yields?

2. Also how many DC needed to be added to see some IL-2/IFN production
by T cells (for 1x10 5 T cells) ?

Thanks for all suggestions.

Mohan
----------------------------------------------------------------------
Mohan N
D1002 DFCI
44 Binney St
Boston MA 02115
mohan_natesan at dfci.harvard.edu



More information about the Immuno mailing list