» Articles » PMID: 20595955

Establishment of Primary Cultures of Human Brain Microvascular Endothelial Cells to Provide an in Vitro Cellular Model of the Blood-brain Barrier

Abstract

We describe a method for generating primary cultures of human brain microvascular endothelial cells (HBMVECs). HBMVECs are derived from microvessels isolated from temporal tissue removed during operative treatment of epilepsy. The tissue is mechanically fragmented and size filtered using polyester meshes. The resulting microvessel fragments are placed onto type I collagen-coated flasks to allow HBMVECs to migrate and proliferate. The overall process takes less than 3 h and does not require specialized equipment or enzymatic processes. HBMVECs are typically cultured for approximately 1 month until confluent. Cultures are highly pure ( approximately 97% endothelial cells; approximately 3% pericytes), are reproducible, and show characteristic brain endothelial markers (von Willebrand factor, glucose transporter-1) and robust expression of tight and adherens junction proteins as well as caveolin-1 and efflux protein P-glycoprotein. Monolayers of HBMVECs show characteristically high transendothelial electric resistance and have proven useful in multiple functional studies for in vitro modeling of the human blood-brain barrier.

Citing Articles

Permeability Benchmarking: Guidelines for Comparing , , and Measurements.

Jorgensen C, Linville R, Galea I, Lambden E, Vogele M, Chen C J Chem Inf Model. 2025; 65(3):1067-1084.

PMID: 39823383 PMC: 11815851. DOI: 10.1021/acs.jcim.4c01815.


Advances in modeling permeability and selectivity of the blood-brain barrier using microfluidics.

Sun J, Song S Microfluid Nanofluidics. 2025; 28(7.

PMID: 39781566 PMC: 11709447. DOI: 10.1007/s10404-024-02741-z.


GDNF and cAMP significantly enhance blood-brain barrier integrity in a humanized tricellular transwell model.

Kanjanasirirat P, Saengsawang W, Ketsawatsomkron P, Asavapanumas N, Borwornpinyo S, Soodvilai S Heliyon. 2024; 10(20):e39343.

PMID: 39492921 PMC: 11530796. DOI: 10.1016/j.heliyon.2024.e39343.


Meta-analysis of the make-up and properties of in vitro models of the healthy and diseased blood-brain barrier.

Shamul J, Wang Z, Gong H, Ou W, White A, Moniz-Garcia D Nat Biomed Eng. 2024; .

PMID: 39304761 DOI: 10.1038/s41551-024-01250-2.


An easy-to-perform method for microvessel isolation and primary brain endothelial cell culture to study Alzheimer's disease.

Chen Y, Huang X, Chen H, Yi C Heliyon. 2024; 10(12):e33077.

PMID: 38994107 PMC: 11238044. DOI: 10.1016/j.heliyon.2024.e33077.


References
1.
Mkrtchyan H, Scheler S, Klein I, Fahr A, Couraud P, Romero I . Molecular cytogenetic characterization of the human cerebral microvessel endothelial cell line hCMEC/D3. Cytogenet Genome Res. 2009; 126(4):313-7. DOI: 10.1159/000253080. View

2.
Bowman C, Witte M, Witte C, Way D, Nagle R, Copeland J . Cystic hygroma reconsidered: hamartoma or neoplasm? Primary culture of an endothelial cell line from a massive cervicomediastinal hygroma with bony lymphangiomatosis. Lymphology. 1984; 17(1):15-22. View

3.
Weksler B, Subileau E, Perriere N, Charneau P, Holloway K, Leveque M . Blood-brain barrier-specific properties of a human adult brain endothelial cell line. FASEB J. 2005; 19(13):1872-4. DOI: 10.1096/fj.04-3458fje. View

4.
Stins M, Gilles F, Kim K . Selective expression of adhesion molecules on human brain microvascular endothelial cells. J Neuroimmunol. 1997; 76(1-2):81-90. DOI: 10.1016/s0165-5728(97)00036-2. View

5.
Zlokovic B . The blood-brain barrier in health and chronic neurodegenerative disorders. Neuron. 2008; 57(2):178-201. DOI: 10.1016/j.neuron.2008.01.003. View