I've begun to establish some PDX lines that consistently passage through mice. At this point, I'd like to make a cell line from these PDX tissues. When I dissociate the tumor tissue and plate it I end up with a lot of mouse stroma that eventually takes over the culture. I'm hoping someone has a good protocol for deriving a purified human tumor line from PDX tissue. We have a Miltenyi MACS system but have not have success using their mouse cell depletion kits after tumor dissociation. At this point any alternative approach could help get over this hurdle.
Hi Steven,
I would try enriching the tumor cells by sorting them (FACS) with human CD44. The HNSCC PDX that I have grown were very positive for this marker.
Best,
Maija
There are mouse cell depletion kits on the market.
This also may give you an idea of what to do.
https://www.jove.com/video/54259/depletion-mouse-cells-from-human-tumor-xenografts-significantly
Good Luck,
Christina
Hello Steven
i have isolated in our lab a murine mammary carcinoma cell line and succeeded in removing the murine fibroblasts using the attached protocol.
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