What does fibronecting coating do? - (Oct/19/2009 )
I am trying to do a primary cell (B cell) culture which requir a feeder layer of irradiated mouse fibroblasts. However I found it's really difficult to harvest the cells without contamination. Will usage of fibronectin-coated culture dishes omit the need of feeder cell layer? Has anybody done primary cell culture using fibronectin-coated plates? Thanks!
-TracyDuke-
TracyDuke on Oct 19 2009, 04:58 PM said:
I am trying to do a primary cell (B cell) culture which requir a feeder layer of irradiated mouse fibroblasts. However I found it's really difficult to harvest the cells without contamination. Will usage of fibronectin-coated culture dishes omit the need of feeder cell layer? Has anybody done primary cell culture using fibronectin-coated plates? Thanks!
The fibronectin will merely adhere the cells to the plates. Feeder cells may provide growth factors, metabolic support, etc. So I guess the answer is it depends on why you're using feeder cells in the first place. ... I'm assuming FACS is out of the question? That'll clean up your cell lines.
As to culturing methods, I'll leave that to people with more cell culture experience than me.
Cheers,
-Carlton
-Carlton H-
Carlton H on Oct 19 2009, 01:23 PM said:
TracyDuke on Oct 19 2009, 04:58 PM said:
I am trying to do a primary cell (B cell) culture which requir a feeder layer of irradiated mouse fibroblasts. However I found it's really difficult to harvest the cells without contamination. Will usage of fibronectin-coated culture dishes omit the need of feeder cell layer? Has anybody done primary cell culture using fibronectin-coated plates? Thanks!
The fibronectin will merely adhere the cells to the plates. Feeder cells may provide growth factors, metabolic support, etc. So I guess the answer is it depends on why you're using feeder cells in the first place. ... I'm assuming FACS is out of the question? That'll clean up your cell lines.
As to culturing methods, I'll leave that to people with more cell culture experience than me.
Cheers,
-Carlton
Thank you Carlton very much for your reply!
I don't think that we need the feeder cells to provide any growth factors in this case becasue we are adding growth factors to the cultrue. FACS sorting is possible too...
-TracyDuke-