I am trying to set up a PD-L1 expressing ovalbumin tumor model (which excludes E.G7 OVA) in my lab and have had trouble with a B16-OVA line ulcerating at seemingly random times, when large but even when very small. We are injecting B6 mice with 500,000 cells that have been rinsed 3x in PBS and are injected subq in 100uL PBS. Has anyone encountered this problem with the B16s? OR does anyone know of another tumor model we could use? (I should note that we received these cells from another lab and are not a reliable vendor, if anyone know of a vendor that sells a B16 OVA that would also be helpful) Thank you!
subset of patients with metastatic melanoma can be successfully treated by the administration of recombinant interleukin-2 (rIL-2), sometimes given together with autologous melanoma-reactive lymphocytes that have been expanded ex vivo (Rosenberg, 1997; Rosenberg, 1999). Recently, a number of different laboratories have used these anti-tumor lymphocytes to clone melanoma-associated antigens, which have generally been nonmutated melanocyte differentiation antigens (MDA), a group that includes MART-1/Melan-A, gp100, tyrosinase, and tyrosinase related proteins (TRP)-1 and -2 (Rosenberg, 1997; Rosenberg, 1999), and others listed in Table 20.1.1. With the possible exception of MART-1/Melan-A, each member of this group of proteins is an enzyme directly involved in the synthesis of eumelanin pigment (Winder et al., 1994; Kameyama et al., 1995).please go to ncbi...details they enormous research material to explain ur concept sir...http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2763508/
Thanks, but I've read that. It states that they ulcerative when large. Nothing about them doing it when small. These B16 also do not express OVA, which I need more than I need a B16. I need an OVA expressing tumor that expresses PD-L1, as my info above states.
You should analyze B16 cell lines from different providers for OVA expression.
However, with my experience, you will encounter this problem of ulceration with any B16 cell lines.
Why do you want a SC model?
You should try the B16F10 metastatic model (injection of the cells into theteil vein). I do not know if it expressing OVA.
Robert
I have used the B16-OVA in C57BL6 - tumors generally ulcerated when > 1.5 cm on bilateral flanks. I used very small TB needles (30 g) and injected very gently into superficial SQ. You might be injecting with too large a needle which causes a tumor seeding too superficially. It also helps to keep the volumes small since too much fluid will put tension on the delicate skin.
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