Does anybody has experience in glioblastoma research and could help me out with an in vitro model to test possible therapeutics on tumor progression etc.?
I saw that people use U-87 cells but I am wondering about the read-out of this system and how to handle positive/negative controls.
Thanks in advance!!
I would be happy to provide some advice, however some more details about what your specific research objectives are would be helpful.
Thank you two for your responses! I will check the paper you attached, definitely sounds interesting. However, for the beginning I think I would like to go with cell lines.
@Pawel: I would like to try different pharmacological agents concerning their ability to stop/decrease GBM progression and/or growth. In order to screen these agents and moreover to get some data necessary for animal work, I would like to use in vitro models. As I said, I am not sure about the read-out of such cell line based models.
OK, well there is a slew of different in vitro experiments you could do on any pharmacological agent on GBM cell lines to test its effect on cell growth, cell survival, cell migration, etc before moving on to an in vivo model. I would suggest doing the in vitro work in more than just U87 cells. Depending on what you observe your drug does to the cells will determine a little but what direction you go in, but direct cell counting, MTT/MTS assays, and IC50 concentration could be a starting point. You would definitely need to use a vehicle as negative control in all of your experiments, but as for a positive control it would depend on what assay you are doing (use various agents or techniques to induce for example, DNA damage - H2AX or TUNEL positivity, cellular senescence - beta gal positivity, apoptosis - cleaved caspase positivity, etc).
If you have an idea of what your drug could be doing, then you would know which assays to focus on at the beginning. Other than that I would suggest looking up a few papers that have done in vitro work in GBM and start from there.
Also, for the sake of future in vivo work do you know whether or not your pharmacological agent is capable of passing through the BBB?
@Julia The World Health Organization classification of brain tumours no longer uses the term "multiforme" and suggests only the use of "glioblastoma" even though the abbreviation used is still GBM.
Thanks again for all your input!
The drug itself is quite probable not to cross the BBB but we are also working on delivery systems for that issue.
I guess we will go for as many in vitro data as possible and try to get in into an in vivo set-up as soon as possible. It's always difficult to predict which amount and which kind of data is sufficient to apply for animal work...
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