Isolation And Culture of Rat Intestinal Mucosal Microvascular Endothelial Cells Using Immunomagnetic Beads And Their Marker Expression

Microvascular endothelial cells (MVECs) have been an important tool in many research fields, while their purification remained challenging. This study aimed to establish a method for isolating rat intestinal mucosal MVECs using immunomagnetic beads, and characterize their proliferation ability and marker expression. Rat jejunum mucosas were scraped and digested by collagenase type II to obtain primary cell culture by the differential adhesion method. Magnetic beads were coated with anti-CD31 antibodies and incubated with the single-cell suspension of the primary cell culture. The CD31 + cells were separated using an automatic magnetic separation system, and their proliferation ability were assayed at different passages. The expression of factor VIII (FVIII), CD31, and CD34 was detected by immunofluorescence staining. Results indicated that rat intestinal mucosal MVECs grew into a contact-inhibited cobblestone-like monolayer after about 6 days, whose proliferation ability had no significant decline at passage 20. Three markers were generally expressed in the cells, and the fluorescence intensity of FVIII was higher after magnetic separation than before, and those of CD31 and CD34 remained similar. In conclusion, highly pure MVECs can be isolated from the primary cell culture of rat intestinal mucosas using magnetic beads coated with anti-CD31 antibodies, and magnetic separation may influence the expression of FVIII.