In vitro culture of cardiomyocytes based on transglutaminase crosslinked gelatin hydrogels
Objective This study aimed to investigate the effect of microbial transglutaminase (mTG) cross-linked gelatin hydrogel on the viability and proliferation of neonatal rat cardiomyocyte (NRCM) and to explore its applicability in myocardial tissue engineering. Methods Gelatin hydrogels crosslinked with microbial transglutaminase were utilized as the substrate for in vitro culture of NRCMs. The growth of cardiomyocytes was observed under a inverted optical phase contrast microscope,and compatibility assessment was performed on cardiomyocytes cultured within one week. Cell viability was evaluated by using live/dead staining (calcein-AM/propidium iodide) to characterize the cellular activity of cardiomyocytes cultured on the gelatin hydrogel surface. Cell proliferation on the hydrogel surface was assessed by using the MTT assay. Immunofluorescence staining with DAPI for cardiac-specific markers cTnI and cTnT was employed to investigate the cardiac origin and maturity of NRCMs cultured on the gelatin hydrogel substrate. Results Gelatin rapidly gelated at 37 ℃ upon the addition of mTG solution. Proliferation of cardiomyocytes and synchronous spontaneous beating behavior were clearly observed under bright-field microscopy over time. Live/dead staining and MTT assay results demonstrated that cardiomyocytes on mTG-GA hydrogels exhibited robust viability and maintained considerable proliferative potential. Immunofluorescence staining for cTnI and cTnT indicated that cardiomyocytes grown on mTG-GA hydrogels not only retained excellent cell viability but also displayed a comparable cardiac cell phenotype and higher expression of characteristic proteins compared to traditional culture on tissue culture plates. Conclusions Microbial transglutaminase-crosslinked gelatin hydrogel exhibits excellent cell compatibility and is a suitable material for cardiomyocyte cultivation, making it conducive to the construction of engineered cardiac tissues and other applications in myocardial tissue engineering.
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维普
