How to isolate cells embedded in Matrigel

Does anyone have successful experience in isolating intact cells embedded in Matrigel for subsequential studies, such as FACS, WB? Thanks in advance!

Hi Doug,

It is very late to respond to you tho I guess this may help others. BD sells dipase to dissolve matrigel and Trevigen may have similar item as well. But I think the use of harvesting solution may affect cell junction molecules as well as surface proteins. For RNA extraction, I saw some people suggests direct application of Trizol onto cells in 3D culture (but never tried). Hope this helps!

-Anna

my reply is also very belated

dispase works well for WB/RNA extraction
it does not work for FACS, the problem is the cells are still too sticky/clog the machine very quickly since the dispase protocol works by centrifugation and you will not get every bit of rBM coating off of the cells
(same for WB, your BCA will be completely useless/you need to quantify empirically)

we have tried many protocols/attempts to get around this problem--to no avail
i know industry sectors are looking into this esp for FACS since i get asked this question at every conference/meeting when i present 3D data!!

good luck!

Hi Cookiedr,

My lab isolated cells using dispase and did culture and FACS analysis with them. Only problem was to have enough number of cells. You may spin down cells at lower rpm/rcf.