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Trans-differentiation of human adipose-derived mesenchymal stem cells into cardiomyocyte-like cells on decellularized bovine myocardial extracellular matrix-based films

  • Yavuz Emre ArslanEmail author
  • Yusuf Furkan Galata
  • Tugba Sezgin Arslan
  • Burak Derkus
Tissue Engineering Constructs and Cell Substrates Original Research
Part of the following topical collections:
  1. Tissue Engineering Constructs and Cell Substrates

Abstract

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.

Notes

Acknowledgements

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.

Supplementary material

10856_2018_6135_MOESM1_ESM.docx (1.2 mb)
Supplementary Materials.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Regenerative Biomaterials Laboratory, Department of Bioengineering, Engineering FacultyCanakkale Onsekiz Mart UniversityCanakkaleTurkey
  2. 2.Department of Biomedical Engineering, Engineering FacultyEskisehir Osmangazi UniversityEskisehirTurkey

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