Isolation and culture of rat microvascular endothelial cells.
The purpose of this study is to identify the separation techniques that result in pure cultures of rat microvascular endothelial cells (MECs). A multistep process is used to optimize the separation of the cells from rat epididymal fat pads, obtaining as pure a culture as possible within a relatively short processing time. The process initially employs the digestion, filtration, and density gradient separation steps. We further describe the use of an attachment phase that allows the differential adherence of contaminating cell types. Immunomagnetic purification is the final step in the process and is performed using anti-PECAM-1 (CD31) monoclonal antibody-labeled DynaBeads.
The Laboratory of Reparative Biology and Bioengineering, Department of Plastic Surgery, University of Texas MD Anderson Cancer Center, Houston, USA.
Cynthia A
CA
Frye
Charles W
CW
Patrick
D000273:Adipose Tissue / Q000098:blood supply; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002469:Cell Separation / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000379:methods; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D008833:Microcirculation; D008858:Microscopy, Phase-Contrast / Q000379:methods; D051381:Rats; D013045:Species Specificity