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Optimization of Microenvironments Inducing Differentiation of Tonsil-Derived Mesenchymal Stem Cells into Endothelial Cell-Like Cells

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

Stem cell engineering is appealing consideration for regenerating damaged endothelial cells (ECs) because stem cells can differentiate into EC-like cells. In this study, we demonstrate that tonsil-derived mesenchymal stem cells (TMSCs) can differentiate into EC-like cells under optimal physiochemical microenvironments.

Methods:

TMSCs were preconditioned with Dulbecco’s Modified Eagle Medium (DMEM) or EC growth medium (EGM) for 4 days and then replating them on Matrigel to observe the formation of a capillary-like network under light microscope. Microarray, quantitative real time polymerase chain reaction, Western blotting and immunofluorescence analyses were used to evaluate the expression of gene and protein of EC-related markers.

Results:

Preconditioning TMSCs in EGM for 4 days and then replating them on Matrigel induced the formation of a capillary-like network in 3 h, but TMSCs preconditioned with DMEM did not form such a network. Genome analyses confirmed that EGM preconditioning significantly affected the expression of genes related to angiogenesis, blood vessel morphogenesis and development, and vascular development. Western blot analyses revealed that EGM preconditioning with gelatin coating induced the expression of endothelial nitric oxide synthase (eNOS), a mature EC-specific marker, as well as phosphorylated Akt at serine 473, a signaling molecule related to eNOS activation. Gelatin-coating during EGM preconditioning further enhanced the stability of the capillary-like network, and also resulted in the network more closely resembled to those observed in human umbilical vein endothelial cells.

Conclusion:

This study suggests that under specific conditions, i.e., EGM preconditioning with gelatin coating for 4 days followed by Matrigel, TMSCs could be a source of generating endothelial cells for treating vascular dysfunction.

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Acknowledgements

This study was supported by the Basic Science Research (NRF-2017M3A9B3063636 and NRF-2017R1A2B4002611), the Bio & Medicial Technology Development Program (NRF-2016M3A9B4919639) and Small Grant for Exploratory Research (NRF-2018R1D1A1A02085696) programs through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning. The paper was also supported by RP-Grant 2019 of Ewha Womans University.

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Author notes

  1. Se-Young Oh and Da Hyeon Choi have contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Medicine, College of Medicine, Ewha Womans University, 260 Gonghang-daero, Gangseo-gu, Seoul, 07804, Republic of Korea

    Se-Young Oh, Yoon Mi Jin, Yeonsil Yu, Ha Yeong Kim, Gyungah Kim & Inho Jo

  2. Ewha Tonsil-derived Mesenchymal Stem Cells Research Center (ETSRC), College of Medicine, Ewha Womans University, 260 Gonghang-daero, Gangseo-gu, Seoul, 07804, Republic of Korea

    Se-Young Oh, Yoon Mi Jin, Yeonsil Yu, Ha Yeong Kim, Gyungah Kim & Inho Jo

  3. School of Biological Sciences, College of Natural Sciences, Chungbuk National University, Chungdae-ro 1, Seowon-Gu, Cheongju, Chungbuk, 28644, Republic of Korea

    Da Hyeon Choi & Yoon Shin Park

  4. Department of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Ewha Womans University, 260 Gonghang-daero, Gangseo-gu, Seoul, 07804, Republic of Korea

    Ha Yeong Kim

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  1. Se-Young Oh
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Correspondence to Yoon Shin Park or Inho Jo.

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The study protocol was approved by the Institutional Review Board of Ewha Womans University Medical Center (IRB No. ECT-11-53-02). Informed consent was confirmed by the IRB.

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Oh, SY., Choi, D.H., Jin, Y.M. et al. Optimization of Microenvironments Inducing Differentiation of Tonsil-Derived Mesenchymal Stem Cells into Endothelial Cell-Like Cells. Tissue Eng Regen Med 16, 631–643 (2019). https://doi.org/10.1007/s13770-019-00221-y

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