Help with Cell attachment problem - (Feb/10/2006 )
I am working with LnCAP cell line. The cells attaches the bottom loosely and grow in clusters. When I do the treatment such as washing it with PBS or changing medium, many cells come off and float. What can I do to solve the problem?
Waiting for your kindly help........
Hi, I worked with LNCaP also, did not have this problem...
I would check that the flasks or dishes you are using are tissue-culture treated... Cells won't attach unless the flasks are tissue-culture treated... It may be easy to overlook (someone ordered wrong or back in the day they did suspension cultures or the flasks are ordered for coating with ECM etc...) so check that first... if they are okay then what media formulation are you using?
Thanks for HTH's reply.
I don't have the attachment problem with flasks but with plates such as 6 well plates or 96 well plates. I guess the pressure on the cells creating by adding or aspirating medium disturbe the loosely attached cells and cause cells to detach.
I am sure that the plates I am using are for tissue-culture. And the culture medium I am using is as the recommendation by ATCC--RPMI 1640 with 10% FBS and 1% pen-step.
LnCAP cells loose attachment easily. We do not even let our LnCAP cells smell trypsin. We subcultures LnCAP cells only by gently shaking the flask to dissociate the cells. After trypsinization this cell lines is difficult to get to reattach