Hello everyone,
I recently started culturing the cells and that since i am new into sterile working and all these, i just wanted to practice a few basics of cell culture and started in absence of growth factor.
Now that I did my subculture for the third time, i see that the cells are having problem with attachment, so want to know if this lack of attachment could in any way be attributed to the absence of growth factors or shall i start looking for other possible reasons.
Hope to get some replies soon
Thanks u all
Growth factors are so important in cell adhesion, so this might be a reasonable explanation. But what type of cell are you culturing? and how did you do you subculture?
Thank you for your response. Yeah I had some thought that growth factors in some way might have caused it but again i wanted to ask in here in this forum as to how long can we go on culturing the cells in absence of any growth factors. This differs in different cell lines I understand, just wanted to know the approx idea about it.
I am culturing OLN-93,the rat OPC cells, subculture by d normal method of PBS washing, trypsinization, inactivating with serum, centrifuging, resuspending, and seeding the cells into new flasks.
@ Walter
I am using 10% FBS in to DMEM and pen-strep combination. Well I have observed the cells under high power and had they been microbial contamination would it not have turned the solution hazy quite in few days time, the solution is absultely clear to naked eyes, though some cells are in suspended form.
I now think it was probably during trypsinization that i left my cells long in trypsin EDTA solution and it may be the negative effect of EDTA on the cells.
Anyways to rule out the contamination should I carry on with microbial tests.
Thank you for your comments
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