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CD105+ Porcine Endometrial Stromal Mesenchymal Stem Cells Possess Differentiation Potential Toward Cardiomyocyte-Like Cells and Insulin-Producing β Cell-Like Cells In Vitro

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Abstract

Porcine mesenchymal stem cells (MSCs) are similar to human MSCs, hence considered a valuable model for assessing potential for cell therapy. Porcine adipose-derived MSCs (AD-MSCs) and endometrial stromal MSCs (EMSCs) displayed fibroblast-like morphology and were positive for MSC markers CD73, CD90, and CD105 and negative for hematopoietic markers CD34 and CD45. The EMSCs had similar or slightly higher growth rate compared to AD-MSCs, and similar percentage of cells of both EMSCs and AD-MSCs were at G0/G1 and G2/M phases; however, EMSCs had significantly (P <.05) higher percentage of cells at S phase of cell cycle than AD-MSCs. Transdifferentiation ability to cardiomyocyte-like cells was confirmed in differentiated cells by the expression of lineage-specific marker genes such as DES, ACTA2, cTnT, and ACTC1 by real-time quantitative polymerase chain reaction (RT-qPCR). Furthermore, cardiomyocyte-specific protein markers cTnT and ACTC1 were expressed in completely differentiated cells. Endodermal differentiation capacity of EMSCs to pancreatic β cell-like cells was evident with the changes in morphology and the expression of β-cell—specific marker genes such as PDX1, GLUT2, SST, NKX6.1, PAX4, and NGN3 as analyzed by RT-qPCR. The differentiated cells secreted insulin and C-peptide upon glucose challenge and also they expressed insulin, PDX1, PAX4, NGN3, and GLUT2 at protein level as assessed by immunostaining confirming the successful differentiation to β cell-like cells. Porcine EMSCs possess all the characteristics of MSCs and are suitable model for studying molecular mechanisms of cellular differentiation.

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Authors and Affiliations

  1. Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501, Jinju-daero, Jinju, 660-701, Republic of Korea

    Raghavendra Baregundi Subbarao PhD, Sharath Belame Shivakumar PhD, Yong-Ho Choe DVM, Young-Bum Son DVM, Hyeon-Jeong Lee DVM, Imran Ullah PhD, Si-Jung Jang DVM, Sung-Lim Lee PhD & Gyu-Jin Rho PhD

  2. Animal Biotechnology Division, National Institute of Animal Science, RDA, Wanju-gun, Jeollabuk-do, Republic of Korea

    Sun-A. Ock PhD

  3. Research Institute of Life Sciences, Gyeongsang National University, Jinju, Republic of Korea

    Gyu-Jin Rho PhD

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  1. Raghavendra Baregundi Subbarao PhD
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  2. Sharath Belame Shivakumar PhD
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Correspondence to Gyu-Jin Rho PhD.

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Authors’ Note

Raghavendra Baregundi Subbarao and Sharath Belame Shivakumar contributed equally to this manuscript.

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Subbarao, R.B., Shivakumar, S.B., Choe, YH. et al. CD105+ Porcine Endometrial Stromal Mesenchymal Stem Cells Possess Differentiation Potential Toward Cardiomyocyte-Like Cells and Insulin-Producing β Cell-Like Cells In Vitro. Reprod. Sci. 26, 669–682 (2019). https://doi.org/10.1177/1933719118786461

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