Construction of cardiomyoblast sheets for cardiac tissue repair: comparison of three different approaches
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Recently, cell sheet engineering has emerged as one of the most accentuated approaches of tissue engineering and cardiac tissue is the pioneering application area of cell sheets with clinical use. In this study, we cultured rat cardiomyoblasts (H9C2 cell line) to obtain cell sheets by using three different approaches; using (1) thermo-responsive tissue culture plates, (2) high cell seeding density/high serum content and (3) ascorbic acid treatment. To compare the outcomes of three methods, morphologic examination, immunofluorescent stainings and live/dead cell assay were performed and the effects of serum concentration and ascorbic acid treatment on cardiac gene expressions were examined. The results showed that cardiomyoblast sheets were successfully obtained in all approaches without losing their integrity and viability. Also, the results of RT-PCR analysis showed that the types of tissue culture surface, cell seeding density, serum concentration and ascorbic acid treatment affect cardiac gene expressions of cells in cell sheets. Although three methods were succeeded, ascorbic acid treatment was found as the most rapid and effective method to obtain cell sheets with cardiac characteristics.
KeywordsH9C2 cell line Cardiac tissue Cell sheet engineering Ascorbic acid Thermo-responsive surface
This study was financially supported by The Hacettepe University Scientific Research Projects Coordination Unit Project No. FBA-2017-16248. The authors would like to thank Selin Gümüşderelioğlu for English editing of text language.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Dergilev KV, Makarevich PI, Tsokolaeva ZI, Boldyreva MA, Beloglazova IB, Zubkova ES, Menshikov MY, Parfyonova YV (2017) Comparison of cardiac stem cell sheets detached by Versene solution and from thermoresponsive dishes reveals similar properties of constructs. Tissue Cell 49:64–71CrossRefGoogle Scholar
- Dergilev K, Tsokolaeva Z, Makarevich P, Beloglazova I, Zubkova E, Boldyreva M, Ratner E, Dyikanov D, Menshikov M, Ovchinnikov A, Ageev F, Parfyonova Y (2018) C-kit cardiac progenitor cell based cell sheet improves vascularization and attenuates cardiac remodeling following myocardial infarction in rats. BioMed Res Int 2018, 3536854. https://doi.org/10.1155/2018/3536854 CrossRefGoogle Scholar
- Ivanyuk D, Budash G, Zheng Y, Gaspar JA, Chaudhari U et al (2015) Ascorbic acid-induced cardiac differentiation of murine pluripotent stem cells: transcriptional profiling and effect of a small molecule synergist of wnt/β-catenin signaling pathway. Cell Physiol Biochem 36:810–830CrossRefGoogle Scholar
- Kato S, Shanley JR, Fox JC (1996) Serum stimulation, cell-cell interactions, and extracellular matrix independently influence smooth muscle cell phenotype in vitro. Am J Pathol 149:687–697Google Scholar
- Ricotti L, Polini A, Genchi GG, Ciofani G, Iandolo D et al (2012) Proliferation and skeletal myotube formation capability of C2C12 and H9c2 cells on isotropic and anisotropic electrospun nanofibrous PHB scaffolds. Biomed Mater 7, 035010. https://doi.org/10.1088/1748-6041/7/3/035010 CrossRefGoogle Scholar