DISSOCIATION MEDIA

 

Hanks solution -modified
5.0 ml
Trypsin (1:250)
0.05 g
gives 0.1% trypsin solution

 

PROCEDURE

1. Collect embryonic tissue (8-24 hippocampi) in Hanks modified solution.

2. Incubate tissue in dissociation media, 5.0 ml for 15 minutes at 37 degrees C.

3. Rinse 3X @ 5 minutes in Hanks modified solution.

4. Pass cells repeatedly through fire-polished Pasteur pipette until no lumps of tissue appear.

5. Count in hemocytometer and assay viability by exclusion of trypan blue.