I am experimenting on chitosan sheets for cell adherence and proliferation studies using NIH3T3 fibroblast cells. I am coating the tissue culture plate with 200ul-1ml of chitosan (1mg/ml) later dried and alcohol sterilized. After seeding the cells, they are not surviving but the control plates survives. Chitosan is known for its cell adherence and proliferation activity, but I don't know why it's not happening in my system. I also tried with chitosan from different vendors. Cells are not adhering to that also.
Kindly help me out on this.
thank you.
I gusse you need to get rid of alcohol completly ..So, after sterlizing with alcohol wash with PBS several time, at least 3 x. ..why you donot use UV-light to sterilization.
Lieder et al. Biological Procedures Online 2013, 15:11
http://www.biologicalproceduresonline.com/content/15/1/11
@ Maryam Hatami
i did PBS washing and also i done that uv treatment procedure . but still, its the same.
What plates do you use, I mean 6-well, 12? 1 mL to cover a well is a lot. Try sterilization with UV w/o alcohol then PBS rinse and dry. I know it may sound crazy but it is possible that the line you use is susceptible to chitosan, simply "does not like" it and this is why does not grow. Other options are to cover plate for ex with collagen and also try to seed other fibroblastic line or/and primary fibroblasts on chitosan. If you need primary fibroblasts for testing I have some which derive from gingiva (control, gingival fibromatosis, periodontitis), and I can share with you.
Good luck! k
... and you use conc of 1mg/mL, right? You can try to use lower concentrations, e.g. 500 ug/mL, 100 ug/mL. ..and do you use PBS as a diluent?
Does that mean . if a cell line doent like to grow ona material then it will not. is it the material fault or the cell lines fault ?.. as far as ive noticed with the chitosan (lower concentration) coated plates there are cells on the areas where chitosan is not coated ..so i think there may be an adhrence problem ...but all literatures says complete opposite fact about chitosan...
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