Synopsis
We previously succeeded in fabricating a microvascular endothelial model with excellent endothelial barrier function by coculturing human microvascular endothelial cells (HMVECs) and human dermal fibroblasts (HDFs) via a collagen vitrigel membrane (CVM). The disadvantage of this model is the low reproducibility especially due to the low proliferation potential of HMVECs. Meanwhile, HMEC-1 cells, a well-known immortalized HMVEC line, show low endothelial barrier function although they have high proliferating potential. This study aimed to create a hybrid cell line by fusing HMVECs and HMEC-1 cells. Hybrid cells were composed of relatively uniform cobblestone-shaped cells compared to HMEC-1 cells and proliferated well. Hybrid cells cultured in a CVM chamber for 14 days highly expressed vascular endothelial (VE)-cadherin at the plasma membrane while VE-cadherin was mainly expressed in the cytoplasm in HMEC-1 cells. The transendothelial electrical resistance (TEER) value of the coculture model of hybrid cells and HDFs via a CVM gradually increased and reached over 30 Ω·cm2 on day 14. On the other hand, the value of the coculture model of HMEC-1 cells and HDFs via a CVM reached a plateau at approximately 10 Ω·cm2. In conclusion, hybrid human microvascular endothelial cells prepared by a cell fusion technique can express high endothelial barrier function and may be useful for quantitative analyses of microvascular permeability.
Key words: microvascular endothelial cells, endothelial barrier, collagen vitrigel membrane, cell fusion technique
