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Effect of Matrigel on formation of cell sheet by co-culture of human induced pluripotent stem cells and polycaprolactone

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Author:
No author available
Journal Title:
Chinese Journal of Biomedical Engineering
Issue:
5
DOI:
10.3760/cma.j.cn115668-20220516-00109
Key Word:
人诱导多能干细胞;聚己内酯;基质胶;细胞补片;细胞增殖;Human induced pluripotent stem cells;Polycaprolactone;Matrigel;Cell sheet;Cell proliferation

Abstract´╝Ü Objective:To investigate the effect of Matrigel on human induced pluripotent stem cells (hiPSCs) adhesion and proliferation on PCL by co-culturing Matrigel-coated polycaprolactone (PCL) with hiPSCs in vitro which simulates the natural extracellular matrix in composition and structure.Methods:After resurgence and culture, the hiPSCs were inoculated onto a Matrigel-coated traditional Petri dish, uncoated PCL, and Matrigel-coated PCL respectively, thereby divided to three groups. After 24 h, the cell growth was examined by 4’, 6-diamidino-2-phenylindole (DAPI) fluorescence under a fluorescence microscope. The stemness of hiPSCs was identified by fluorescence of OCT4, a hiPSCs stemness marker. After fixation, scanning electron microscopy was performed to observe the surface morphology of the cell sheet. The cell viability was determined by CCK-8 assay on days 1, 3 and 5 of culture. The cell growth and proliferation were studied using growth curve.Results:OCT4 fluorescent assay showed green fluorescence emitting from all the three groups, indicating that hiPSCs remained stemness after formation of cell sheets. DAPI fluorescent assay showed blue fluorescence emitting from all the three groups, and that the cells presented clone-like growth with uniform morphology. Scanning electron microscopy showed that the hiPSCs adhered to and grew on Matrigel-coated PCL with clearly visible cell contours and normal morphology. The cell viability in the Matrigel-coated PCL group was greater than that in uncoated PCL group on days 1 and 3 of co-culture (1.905±0.080 vs 1.657±0.027, 2.375±0.096 vs 2.054±0.078, both P<0.05) , but there was no significant difference between the Matrigel-coated and uncoated PCL group on day 5 of co-culture. The cell viability in either Matrigel-coated or uncoated PCL group was greater than that in the Matrigel-coated traditional Petri dish group on days 1, 3 and 5. As shown by growth curve, the cell viability in Matrigel-coated PCL group reached its plateau on day 3 with no statistical difference from that on day 5, and was close to the day-5 absorbance ( A value) in the uncoated PCL group. In the uncoated PCL group, the cell viability was greater on day 5 than that on day 3 ( P<0.05) . These findings indicated that the proliferation of hiPSCs was the most rapid in Matrigel-coated PCL group. Conclusion:Co-culturing hiPSCs with Matrigel-coated or uncoated PCL can lead to formation of cell sheets. Matrigel-coated PCL can offer a better environment for cell growth, adhesion and proliferation. Co-culturing hiPSCs with PCL significantly promotes formation of cell sheets.

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