Three‐dimensional collagenous niche and azacytidine selectively promote time‐dependent cardiomyogenesis from human bone marrow‐derived MSC spheroids

Jyotsna Joshi; Gautam Mahajan; Chandrasekhar Kothapalli

doi:10.1002/bit.26714

journal article Apr 27, 2018

Three‐dimensional collagenous niche and azacytidine selectively promote time‐dependent cardiomyogenesis from human bone marrow‐derived MSC spheroids

Jyotsna Joshi Gautam Mahajan Chandrasekhar Kothapalli

Biotechnology and Bioengineering Vol. 115 No. 8 pp. 2013-2026 · Wiley

View at Publisher Save 10.1002/bit.26714

Abstract

AbstractEndogenous adult cardiac regenerative machinery is not capable of replacing the lost cells following myocardial infarction, often leading to permanent alterations in structure‐function‐mechanical properties. Regenerative therapies based on delivering autologous stem cells within an appropriate 3D milieu could meet such demand, by enabling homing and directed differentiation of the transplanted cells into lost specialized cell populations. Since type I collagen is the predominant cardiac tissue matrix protein, we here optimized the 3D niche which could promote time‐dependent evolution of cardiomyogenesis from human bone marrow‐derived mesenchymal stem cells (BM‐MSC). 3D collagen gel physical and mechanical characteristics were assessed using SEM and AFM, respectively, while the standalone and combined effects of collagen concentration, culture duration, and 5‐azacytidine (aza) dose on the phenotype and genotype of MSC spheroids were quantified using immunofluorescence labeling and RT‐PCR analysis. Increasing collagen concentration led to a significant increase in Young's modulus (p < 0.01) but simultaneous decrease in the mean pore size, resulting in stiffer gels. Spheroid formation significantly modulated MSC differentiation and genotype, mostly due to better cell–cell interactions. Among the aza dosages tested, 10 μM appears to be optimal, while 3 mg/ml gels resulted in significantly lower cell viability compared to 1 or 2 mg/ml gels. Stiffer gels (2 and 3 mg/ml) and exposure to 10 μM aza upregulated early and late cardiac marker expressions in a time‐dependent fashion. On the other hand, cell–cell signaling within the MSC spheroids seem to have a strong role in influencing mature cardiac markers expression, since neither aza nor gel stiffness seem to significantly improve their expression. Western blot analysis suggested that canonical Wnt/β‐catenin signaling pathway might be primarily mediating the observed benefits of aza on cardiac differentiation of MSC spheroids. In conclusion, 2 mg/ml collagen and 10 μM aza appears to offer optimal 3D microenvironment in terms of cell viability and time‐dependent evolution of cardiomyogenesis from human BM‐MSCs, with significant applications in cardiac tissue engineering and stem cell transplantation for regenerating lost cardiac tissue.

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Metrics

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Citations

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References

Details

Published: Apr 27, 2018
Vol/Issue: 115(8)
Pages: 2013-2026
License: View

Authors

J

Jyotsna Joshi

Department of Chemical and Biomedical Engineering Cleveland State University Cleveland Ohio

G

Gautam Mahajan

Department of Chemical and Biomedical Engineering Cleveland State University Cleveland Ohio

C

Chandrasekhar Kothapalli

Department of Chemical and Biomedical Engineering Cleveland State University Cleveland Ohio

Funding

NSF-DMR Award: 1126126

NSF-CBET Award: 1337859

Cellular and Molecular Medicine Specialization fellowship at CSU

Cite This Article

Jyotsna Joshi, Gautam Mahajan, Chandrasekhar Kothapalli (2018). Three‐dimensional collagenous niche and azacytidine selectively promote time‐dependent cardiomyogenesis from human bone marrow‐derived MSC spheroids. Biotechnology and Bioengineering, 115(8), 2013-2026. https://doi.org/10.1002/bit.26714

Three‐dimensional collagenous niche and azacytidine selectively promote time‐dependent cardiomyogenesis from human bone marrow‐derived MSC spheroids

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