The establishment and identification of human umbilical artery endothelial cells-smooth muscle cells co-culture model / 复旦学报（医学版）

ABSTRACT

Objective To establish a human umbilical artery EC-SMC co-culture model, and mimic the morphological and functional characteristics of human arterial wall, for further reseach of the pathological mechanism and therapy of atherosclerosis and imflammatory damage. Methods We secceeded in the primary culture of human umbilical artery endothelial cells (HUAEC) and human umbilical artery smooth muscle cells (HUASMC) by collagenase perfusion digestion and tissue planting, respectively. HUASMCs were incubated in a medium with ascorbic acid at the concentration greater than 50 μg/mL to produce collagen, which was considered as the extracellular matrix for ECs. Then HUAECs were seeded directly upon HUASMCs in a saturate density for sufficient direct physical interaction between ECs and SMCs. The morphological characteristic of EC-SMC co-culture was identified by immunofluorescence staining, and the function of EC-SMC co-culture was identified by Dil-Ac-LDL uptake test. Results The morphological identification showed that the entire surface of HUASMCs was covered by a confluent monolayer confluent monolayer, which indicated that the model had simulated the morphological characteristic of human arterial wall. The results of Dil-Ac-LDL uptake test showed that there was a fluorescent signal in HUAECs. Compared with EC monoculture, the Dil-Ac-LDL uptake of HUAECs was increased significantly in the co-culture system. All the reseach results indicated that there was an interaction between HUAECs and HUASMCs in the co-culture system. Conclusions In the present study, human umbilical artery EC-SMC co-culture model was constructed successfully, which could mimic the morphological characteristic and basic functions of human arterial wall.