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Fabrication of Prevascularized Cell Sheet
Ren Liling1*, Liu Bin1, Ma Dongyang2, Feng Yuxia1, Chen Jia1
1School of Stomatology, Lanzhou University, Lanzhou 730000, China; 2Department of Oral and Maxillofacial Surgery,Lanzhou General Hospital, Lanzhou Command of Chinese People’ s Liberation Army, Lanzhou 730052, China
Abstract: The aim was to explore a new method to acquire capillary-like networks in vitro to realize vascularized engineered tissues. Human mesenchymal stem cells (hMSCs) were cultured on a cell culture dish at a cell density of 9×104/cm2 to form a thick cell sheet. Human umbilical vein endothelial cells (HUVECs) were seeded onto the surface of hMSCs sheet at a cell density of 5×104/cm2. At designated time points， the image of microscope showed that HUVECs migrated and aligned arrangement on the hMSCs cell sheet. This migration of HUVECs made the hMSCs cell sheet matrix rearrange， leading to the formation of aligned microgrooves and intracellular vacuoles. The immunofluorescent images for CD31 revealed a progressive process of lumen morphogenesis. Immunofluorescent staining for CD90 showed an aligned arrangement around the HUVECs， which suggested that hMSCs supported the growth of HUVECs as pericytes. Immunofluorescent staining for α-SMA indicated that few expressions of α-SMA were observed after 10 days， which suggested that hMSCs have low ability of differentiating into myoblast under these current experimental conditions. These results indicate that it is feasible for endothelial cells to assemble into a 3-dimensional prevascular network in hMSCs cell sheet. Our prevascularizing method using coculture model of culturing endothelial cells onto a hMSCs sheet provides a substantial advance for developingvarious types of 3-dimensional tissues and contributes to regenerative medicine.
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