A Modified Protocol for the Isolation, Culture, and Characterization of Human Smooth Muscle Cells from the Umbilical Cord

• Published in: Methods and protocols 2023-05, Vol.6 (3), p.54
• Main Authors: Ibrahim, Asmaa Q, Abdullah, Mohammed S, Ahram, Mamoun, Abdalla, Shtaywy
• Format: Article
• Language: English
• Subjects: Analysis; Arteriosclerosis; Atherosclerosis; Cardiovascular diseases; Cell culture; Cells; Collagen; Connective tissue; Cytology; Dissection; Endothelial cells; Enzymes; Fibroblasts; Gene expression; Genotype & phenotype; Health aspects; isolation protocol; Morphology; Penicillin; Protocol; Reverse transcription; Smooth muscle; Umbilical cord; Umbilical vein; vascular smooth muscle cells; Veins & arteries; VSMC characterization
• ISSN: 2409-9279; 2409-9279
• DOI: 10.3390/mps6030054

Vascular smooth muscle cells (VSMCs) and vascular endothelial cells are key participants in the pathogenesis of atherosclerosis. Human umbilical vein endothelial cells (HUVECs) and VSMCs are useful models to design therapeutic strategies for many cardiovascular diseases (CVDs). However, procuring a VSMC cell line by researchers, to model atherosclerosis, for example, is impeded by time and cost limitations, as well as by many other logistic problems in many countries. This article describes a protocol for the quick and cheap isolation of VSMCs from human umbilical cords using a mechanical and enzymatic method. This VSMC protocol yields a confluent primary culture that could be obtained within 10 days and sub-cultured for 8-10 passages. The isolated cells are characterized by their morphology and the expression of mRNA of marker proteins analyzed by reverse transcription polymerase chain reaction (RT-qPCR). The protocol described herein for the isolation of VSMCs from human umbilical cords is easy and is time- and cost-efficient. Isolated cells are useful models for understanding the mechanisms underlying many pathophysiological conditions.