Study On PHBHHx Co-cultured With Mouse Induced Pluripotent Stem Cells For Use As Myocardial Patches In Vitro

Objective: To study the effect of this new type of polymer material on adhesion, proliferation and differentiation of mouse induced pluripotent stem cells via co-culturing in vitro, to find a suitable polymeric biomaterials for mi PSCs attachment, proliferation and differentiation, forming myocardial patches as a new repair method for myocardial infarction, cerebral infarction, as well as cartilage injury such as a variety of tissue damage diseases.Methods: miPSCs were recovered, passaged and cultured with PHBHHx two-dimension films and PHBHHx three-dimension films(membrane perforated, imitate the three dimensional space) together. The morphology of miPSCs attached on the films was visualized under scanning electron microscope(SEM). The cell vitality was detected by CCK-8. DAPI fluorescence staining to observe cell morphology. The mouse induced pluripotent stem cells were induced by differentiation medium that containing vitamin C. Control group did not add any inducer. The survival and differentiation of miPSCs were observed through immunofluorescence and flow cytometry.Results: miPSCs can adhere and survive on PHBHHx films with normal shape. The cell vitality was detected by CCK-8: PHBHHx three-dimensional film(0.836±0.038)> PHBHHx two-dimensional film(0.617±0.019)> traditional cultivation(0.312±0.004)(p<0.05).Vitamin C, as a favourable inducer, can markedly improve the efficiency of mi PSCs differentiation of cardiomyocytes on PHBHHx films. Immunofluorescence results demonstrated positive cTnT expression. Flow cytometry measured cTnT expression: Vitamin C inducer group(47.54%±1.46%)> without any inducer group(7.02%±0.95%)(p<0.05). In the process of miPSCs differentiating into myocardial cell, after adding Vitamin C, the expression quantity of cTnT: PHBHHx three-dimensional films(60.32%±1.76%)> PHBHHx two-dimensional films( 53.31%±1.41%) >traditional cell culture(47.54%±1.46%)(p<0.05).The expression of a-actinin test by flow cytometry got the same result.Conclusions: 1、MiPSCs have good histocompatibility with PHBHHx film and can adhere, survive and differentiate on the PHBHHx film.2、Vitamin C, as a favourable inducer, can markedly improve the efficiency of miPSCs differentiation of cardiomyocytes.3、Relative to PHBHHx two-dimensional film culture and traditional culture, the three-dimensional PHBHHx film culture has a great advantage in the process of miPSCs differentiating into myocardial cells. PHBHHx films have potential to be applied as a myocardial patch for the regeneration of cardiomyocytes in treating myocardial infarction.