Trans-differentiation of human adipose-derived mesenchymal stem cells into cardiomyocyte-like cells on decellularized bovine myocardial extracellular matrix-based films
In this study, we aimed at fabricating decellularized bovine myocardial extracellular matrix-based films (dMEbF) for cardiac tissue engineering (CTE). The decellularization process was carried out utilizing four consecutive stages including hypotonic treatment, detergent treatment, enzymatic digestion and decontamination, respectively. In order to fabricate the dMEbF, dBM were digested with pepsin and gelation process was conducted. dMEbF were then crosslinked with N-hydroxysuccinimide/1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide (NHS/EDC) to increase their durability. Nuclear contents of native BM and decellularized BM (dBM) tissues were determined with DNA content analysis and agarose-gel electrophoresis. Cell viability on dMEbF for 3rd, 7th, and 14th days was assessed by MTT assay. Cell attachment on dMEbF was also studied by scanning electron microscopy. Trans-differentiation capacity of human adipose-derived mesenchymal stem cells (hAMSCs) into cardiomyocyte-like cells on dMEbF were also evaluated by histochemical and immunohistochemical analyses. DNA contents for native and dBM were, respectively, found as 886.11 ± 164.85 and 47.66 ± 0.09 ng/mg dry weight, indicating a successful decellularization process. The results of glycosaminoglycan and hydroxyproline assay, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), performed in order to characterize the extracellular matrix (ECM) composition of native and dBM tissue, showed that the BM matrix was not damaged during the proposed method. Lastly, regarding the histological study, dMEbF not only mimics native ECM, but also induces the stem cells into cardiomyocyte-like cells phenotype which brings it the potential of use in CTE.
Work on this paper was financially supported by Ministry of Science, Industry and Technology, Republic of Turkey (Project ID. 0089.TGSD.2013) and the Scientific and Technological Research Council of Turkey (Project ID. 114S851). We wish to thank Canakkale Onsekiz Mart University, Science and Technology Application & Research Center and MER-TER Medical for collaborating with analyses.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 1.Wang B, Borazjani A, Tahai M, De Jongh Curry AL, Simionescu DT, Guan J, et al. Fabrication of cardiac patch with decellularized porcine myocardial scaffold and bone marrow mononuclear cells. J Biomed Mater Res Part A. 2010;94:1100–10.Google Scholar
- 12.Arslan YE, Hiz MM, Sezgin Arslan T. The use of decellularized animal tissues in regenerative therapies. Kafkas Univ Vet Fak Derg. 2015;21:139–45.Google Scholar
- 15.Arslan YE, Sezgin Arslan T, Derkus B, Emregul E, Emregul KC. Fabrication of human hair keratin/jellyfish collagen/eggshell-derived hydroxyapatite osteoinductive biocomposite scaffolds for bone tissue engineering: From waste to regenerative medicine products. Colloids Surf B Biointerfaces. 2017;154:160–70.CrossRefGoogle Scholar
- 21.Gilbert TW, Sellaro TL, Badylak SF. Decellularization of tissues and organs. Biomaterials. 2006;27:3675–83.Google Scholar
- 25.Poornejad N, Nielsen JJ, Morris RJ, Gassman JR, Reynolds PR, Roeder BL, et al. Comparison of four decontamination treatments on porcine renal decellularized extracellular matrix structure, composition, and support of human renal cortical tubular epithelium cells. J Biomater Appl. 2016;30:1154–67.CrossRefGoogle Scholar
- 34.Bonvillain RW, Scarritt ME, Pashos NC, Mayeux JP, Meshberger CL, Betancourt AM, et al. Non human primate lung decellularization and recellularization using a specialized large-organ bioreactor. J Vis Exp. 2012;18:2437–52.Google Scholar