10-20 thousand GL261 in 1-2 microlitre will make GBM model in mice. This cell line is from mouse GBM and can be implanted in immunocompetent mouse.

If you want to have human GMB model in immunocompromised mouse then use U87 or U251. However, GMB cells from explanted tumor samples will make better clinically relevant GBM. Find out a collaborator who can send you GBM cancer stem cell line cells.

In mouse you can inject 1.5 to 2 microlitre orthotopically. For human cells you may need to injected large number of cells ~100k.

Hi,

To develop an orthotopic glioblastoma mouse model, you will require to perform an intracranial injection using a stereotaxic setup. You need to mark the location 2mm below bregma and drill a hole through the skull.

NOTE: The drilling is a crucial step of this process. Be gentle yet firm. Drill only till you see the white powder from the skull coming out. 

For injection, you will require a stereotaxic needle (Hamilton syringe). You need to inject at a depth of 3mm into the brain. You can inject up to 2x10^5 cells in 5ul volume and can use U87, U251, and U118 cell lines. The time taken for the tumor to develop may depend on several factors such as the health of your cells, injection at the correct site, the condition of your mouse, etc. If all goes well, it shouldn't take more than 3-4 weeks max to have a nice spread of tumor ready.

Good luck,

Amir

Thanks a lot to Ali and Amirali  for giving a valuable tips

Dear Ali & Amirali,

Please can you tell me the coordinates ( x, y and z from bregma ) to inject the cells for orthotopic GBM Model development using U87 cell line.

Here attached is an article with the GL261 model (Verschuere et al., Int J Cancer 2013).

We routinely use the Hs683 GBM model (but with oligodendroglial origin) because it mimic temozolomide response as in clinics (see the attached article by Le Mercier M et al., JNEN 2008).

The U87 model develops as a compact mass and does definitively not mimic the clinical situation in terms of parenchyme invasion (see Camby et al., JNEN 2002). In contrast, it isan interesting model to stury tumor neoangiogenesis (Lamour et al. Int J Cancer 2010).

Other GBM orthothotopic xenografts are described in Branle et al. Cancer 2002, and Belot et al. 2001.

Best regards

Robert