Thank you! Will check it out....

Hi Claretta,

I used to do this back in the day. Here's a pretty good protocol from one of our papers. This protocol is for rat but mouse brain will be the same.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1385088/?page=2

Most recent answers should be on intechopen.com

Hi,

I suggest you to look this paper, I found it really well described.

A simple method for isolation and characterization of mouse brain microvascular endothelial cells

Zhenhua Wu a, Florence M. Hofman b, Berislav V. Zlokovic a,*

Journal of Neuroscience Methods 130 (2003) 53-63

Sapatino, B., Welsh, C.J.R., Smith, C.A., Bebo, B. and Linthicum, D.S. (1993) Cloned mouse cerebrovascular endothelial cells that maintain their differentiation markers for Factor VIII, low density lipoprotein and angiotensin-converting enzyme. In Vitro Cell. Dev. Biol. 29A, 923-928.

Hi Claretta, we have just published a rat brain microvessel protocol that is extremely easy, robust and gives high yields. it will be equally applicable to mouse. Watson et al, 2013 BMC Neuroscience. Good luck!

Hi Claretta--You don't mention whether you are isolating brain microvessels to dissociate the endothelial cells for culture, or to perform studies on the intact(ish) microvessels themselves. If you are interested in the latter, David Miller's protocol originally described here:

http://molpharm.aspetjournals.org/content/58/6/1357.long

(and in many subsequent publications of his) produces the cleanest preps with the highest yields among everything I've tried. It also has the advantage of producing metabolically active microvessels. It is, however, fairly labor intensive to do correctly (one person working alone would probably spend the better part of a workday isolating vessels from 10 mouse brains, for example.)

If you are less concerned about purity of the prep, but still want intact vessels rather than dissociated cells for culture, the protocol we used here (described in section 2.5):

http://www.sciencedirect.com/science/article/pii/S0006899304013575

is much, much faster, very easy to perform, and yields a lot of vessels. But it does tend to contain a lot of debris in the final pellet.

Good luck!

Thanks for your answer Brian. I am looking to start with working on fairly intact microvessels and then based on the results I might or might not need to work with dissociated cell in culture. This forum has given me so many options, I don't know where to start but I am plowing through it one by one. I really appreciate all the help from everyone here!

Old topic but....

@Claretta

Did you find a way finally?

@Brian

When you applied David Miller's protocol, was it on mouse brain ?

Because people tend to think that what's done on other animal models is also applicable on mouse but it is really not the case : the mouse brain is very small and the yield is so low. And accoding to the protocole they used 200um nylon mesh, which differs from the methods I found where they use 100um meshes and below.

So if both of you could share your insights on this, it would really be appreciated.

Thanks