I am trying to culture glioblastoma cells from biopsies (5-6 mm) taken from GBM patients. The tumor : background parenchyma ratio will be variable. Anyone with experience on culture conditions:
1) Tissue dissociation; enzymatic vs mechanical? Protocols? Culture conditions: Adherent vs free-floating spheroids? If adherent, tissue culture treated vs non tissue culture treated flasks? Astrocyte co-culture or others?
2) Culture media; defined vs serum based? Growth factors EGF, FGF, PDGF, IGF? Concentrations? Glucose, Pyruvate,...etc concentrations? pH? O2 and CO2 %?
3) Optimal passage time for propagation?
Any help is much appreciated.
Thanks
Osama
Hey Osama, maybe this article can help.
In any case passage time varies depending on each patients cells characteristics, and unfortunately it also changes as you keep passaging them. It's better to go by confluency. We used laminin, but it's tricky.
S. M. Pollard, K. Yoshikawa, I. D. Clarke, D. Danovi, S. Stricker, R. Russell, J. Bayani, R. Head, M. Lee, M. Bernstein, J. A. Squire, A. Smith, and P. Dirks, “Glioma Stem Cell Lines Expanded in Adherent Culture Have Tumor-Specific Phenotypes and Are Suitable for Chemical and Genetic Screens,” Cell Stem Cell, vol. 4, no. 6, pp. 568–580, Oct. 2009
Hey Virginia! Thanks for the response. This is really helpful. Do you know what is the success rate of culture?
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