p****z
发帖数: 31 | 1 问题1: 有人做过H1299的soft agar的growth 吗?用的是0.5% base agar+0.4% top
agar. 看paper上,都能做出来.我做了两次,都没有成功.实验方法与操作应该都没有问
题,同时做的H460长得很好.
问题2:soft agar上长出来的cell colony是cancer stem cell/tumor initiating cell
吗?
谢谢! | h********n
发帖数: 4079 | | H*****e
发帖数: 120 | 3 1. H1299 needs 2-3 weeks for seeing colonies, whereas H460 colonies can be
seen in 5-7 day. If it still does not work, change the mediumn every 4 days
or order new cells from ATCC. We routinely do it and no problem found.
2.It has nothing to do with stem cells. Most stem cell medium are serum
free. But Soft agar uses 10-20% serum (I even saw people use 30% serum for
weak oncogene transformed 3T3.).
Good luck! | p****z
发帖数: 31 | 4 Thanks you very much!
I have several other questions:
Why only a minority of the cancer cells can form colonies in the soft agar?
like 1 or 2 cells per 1000 cells. Is there any characteristic of those soft-
agar-growing cells compared to that of the non-soft-agar growthing
population?
Thanks!
days
for
【在 H*****e 的大作中提到】
: 1. H1299 needs 2-3 weeks for seeing colonies, whereas H460 colonies can be
: seen in 5-7 day. If it still does not work, change the mediumn every 4 days
: or order new cells from ATCC. We routinely do it and no problem found.
: 2.It has nothing to do with stem cells. Most stem cell medium are serum
: free. But Soft agar uses 10-20% serum (I even saw people use 30% serum for
: weak oncogene transformed 3T3.).
: Good luck!
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