Can the 4T1 metastatic breast cancer model - which is a syngeneic xenograft model based on 4T1-12B - be used as a model to study the anticancer effects of plant based compounds without injecting into a balb/c mice? Can one culture it directly in flasks, apply plant based extracts and study the effects of plants on the cells?
They can be cultured in a Flask no problem, or a 6, 12, 24, 96 well plate. they stick to anything, grow in a few media
Hi Stephen,
Adrian is correct, you can grow them in flasks very easily. We use filtered/vented T75 flasks and RPMI 1640 media, supplemented with 10% gamma irradiated fetal bovine serum and 1% pen/strep. For drug treatments, we'll use serum-free or 1% serum media. The cells grow quickly- I split the cells 1:10 twice per week. Best of luck with the experiments!
Thanks Ryan And Adrian.
Ryan, do you apply drugs directly to the cells in the flask and then monitor anticancer effect? Can one harvest cells directly from the flask for rna extraction?
Stephen,
We do apply drugs directly to the flask- I count out the required number of cells, and allow them to adhere to the plate overnight. The following day I aspirate the media and replace it with drug-containing media. Our lab usually doesn't use anti-cancer agents, though colleagues of ours do add them to the flask/96-well plate according to their chosen administration schedule. I'm afraid I don't do RNA extraction, sorry!
I recommend something like a taqman cells to ct kit. End point price is comparable to standard RNA extraction, RT, and qPCR. And you can get enough RNA from a single 96 well worth of cells to do really nice PCR
Stephen,
you're talking about two different things.
Yes, you can grow 4T1 cells in culture and study their response to a number of different compounds. You can even screen those compounds to find the active ones. What would be your readout - be it RNA levels, cell viability, cell adhesion etc. depends on your supposed target(s) and your compounds. This is in vitro testing.
However, injecting those cells in mice will create a different story - in vivo testing. And there you may have different results. Especially if you will be looking at major tumor vs metastases. It may or may not correlate with your in vitro data. Unfortunately, life is not easy and life science is not easy :-) Otherwise, we would discover many-many more anti-cancer reagents.
Good luck!
Yes you can. Grow the cells in 24 well plates and test your drug for their efficacy.
Usually it is best to perform in vitro studies of drugs to see their cytotoxicity on a given cell line, translate that concentration into a dosage for in vivo xenograft studies, and use xenografts with the same cell line. In context of 4T1, it has been done many times, and we've run xenografts with it (http://altogenlabs.com/xenograft-models/breast-cancer-xenograft/4t1-xenograft-model/) having a pre-determined concentration of drug/therapeutic along with in vitro results to compare with.
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