Hello:
According to a server my original post did not get
through. My apologies if you are seeing this note a
second time.
We've recently run into a problem with growing
hybridomas (NS-1 / Balb/c). The problem arose during
HAT selection of hybridomas after a fusion. No
hybridomas were left alive after a week. We tested our
media components by growing an established
hybridoma cell line (also NS-l / Balb/c). Cells were
thawed from liquid nitrogen and grown on 10% FCS
/DMEM and tranfered to plates containing:
1) DMEM / 10% FCS
2) DMEM / 10% FCS / HT
3) DMEM / 10% FCS / HAT
4) DMEM / 10% FCS / Aminopterin
Result after 48 hours:
1) Grew well, cells healthy
2) Cells dead/dying
3) Cells dead/dying
4) Cells dead/dying
"Dead" cells appeared rough, with non-uniform size
and were of low cell density. DMEM and HT stocks were
freshly made and the experiment was replicated with
two different lots of FCS. Any suggestions as to why the
cells did not grow in HT or HAT? Thanks in advance.
Les.
Les Willis
Agriculture Canada
Summerland, B.C.
V0H 1Z0
WillisL at em.agr.ca
