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[aegreen]

Has anyone had any experience with primary human skin fibroblasts?

I am finding they grow very slowly and have low protein yield unless they are kept at high confluency. These are an old cell line that I have inherited and I can't find what confluency is recommended.

Any suggestions for confluency?

[Salvador]

Hi aegreen,

It has been a while ago for me, but I never grew them more than 80% confluent, they will otherwise contact inhibit each other and reduce growth rate. You should not split them harshly, it takes them a long time to recover. What do you mean by old? Old as in, have been cultured and passaged for a very long time or are they an old frozen vial from cryo but low passage? If they are late passage they will slow down.
have you considered trying to change media conditions? They like relatively high pH. Replenish the media often.

Best,

S