LNCaP xenografts - need a better protocol (Jul/01/2009 )

Dear Bioforum geniuses,



I need help with LNCaP xenografts. To date, I have done more than 40 xenograft injections of this prostate carcinoma cell line into male athymic homozygous nude mice, and have failed to have my results match those of papers I've read.

Various papers have reported that a per mouse injection of 1-3 million LNCaP cells in a 1:1 mixture with BD Matrigel yield a tumor ~200 mm^3 within 2-3 weeks. I have done injections under the same conditions--but for cell number: I have previously injected 500k LNCaPs/mouse, 5 million LNCaPs/mouse, and 2 million LNCaPs/mouse-- and have not had any growth of a tumor at all until 3-4 weeks after injection.

I had found out that LNCaPs tend to change morphology after passage 10-12, and the first two sets of injections were of passages ~P15-P20. However, I got an vial of LNCaP's at an earlier passage number, and a 2 million LNCaPs/mouse injection of this younger line (passage 6) has (as of week 2+) yielded absolutely nothing in the way of a tumor.

Has anyone here had any experience with LNCaP xenografts, specifically in having tumors grow within 2-3 weeks of injection?

Many thanks,

--Buffonidae

i suffer the same thing....but i try to do xeno with LM2 cell in balb/c
the better that i got after 3 moths testing all protocols was : first amplified your cells and then frezzed them
second: defrost and amplified litte and take care that you shoud to inject early passage.
good luck

Dear Buffonidae,

Have you been able to solve your problems with growing LNCaP xenograft? I am in the same situation and would like some tips.

Thanks

Hola,I´m not an expert and made xenografts few monts ago, but I had realized that my tumoral lines( I haven´t your prostatic LM2) grow faster and better in NOD Scid mice than NUDE, but we have some lines wich grow slowly and bad in both lines too. Why don´t you try inject three or four NOD SCID mice and see if the cells made tumor?, If yes, you could amplify the line in more mice from cells or from the new tumors. I´would sorry that this can´t help you. Buena suerte