Advertisement

Differentiation potential of different regions-derived same donor human Wharton’s jelly mesenchymal stem cells into functional smooth muscle-like cells

  • Dinesh Bharti
  • Sharath Belame Shivakumar
  • Young-Bum Son
  • Young-Ho Choi
  • Imran Ullah
  • Hyeon-Jeong Lee
  • Eun-Jin Kim
  • Sun-A Ock
  • Ji-Eun Park
  • Ji-Kwon Park
  • Dawon Kang
  • Sung-Lim Lee
  • Bong-Wook Park
  • Gyu-Jin RhoEmail author
Regular Article

Abstract

The present study evaluates the transdifferentiation potential of different region-derived same donor Wharton’s jelly MSCs (WJMSCs) into functional smooth muscle-like cells (SMLCs). All regions showed baseline expression for early smooth muscle cell (SMC) markers (αSMA and SM22-α) whereas mid marker CALPONIN gradually reduced during in vitro culture expansion and late marker myosin heavy chain type-11 (MHY-11) was completely absent. Furthermore, WJMSCs were induced to SMLCs using DMEM containing 10% FBS supplemented with different concentrations/combinations of TGF-β1 and PDGF-BB under normoxia (20% O2) condition. Three treatment groups namely group A: 2.5 ng/ml TGF-β1, group B: 5 ng/ml PDGF-BB and group C: 2.5 ng/ml TGF-β1 + 5 ng/ml PDGF-BB were used for the induction of WJMSCs into SMLCs. Cells were evaluated for SMC-specific marker expression at different time intervals. Finally, selection of the SMC-specific highly potent region along with the most suitable treatment group was done on the basis of highest outcome in terms of SMC-specific marker expression and functional competence of transdifferentiated cells. Among all regions, baby region-derived WJMSCs (B-WJMSCs) exhibited highest SMC marker expression and functional ability. To mimic the in vivo physiological conditions, hypoxic conditions (3% O2) were used to evaluate the effect of low oxygen on the SMLC differentiation potential of selected WJMSCs using previously used same parameters. Annexin-V assay was performed to check the effect of cytokines and different oxygen concentrations, which revealed no significant differences. It was concluded that different induction conditions have different but positive effects on the functional SMLC differentiation ability of WJMSCs.

Keywords

Transdifferentiation Smooth muscle cells Mesenchymal stem cells Wharton’s jelly Electrophysiology 

Abbreviations

WJMSCs

Wharton’s jelly mesenchymal stem cells

SMLCs

smooth muscle-like cells

SMC

smooth muscle cell

α-SMA

alpha smooth muscle actin

SM22-α

alpha-smooth muscle 22

MHY-11

myosin heavy chain 11

Notes

Acknowledgements

We are highly thankful to Professor BW Park, Doctor Ji-Eun Park and Doctor Ji-Kwon Park for providing valuable human whole umbilical cord samples. We also acknowledge Professor Dawon Kang for helping us with the electrophysiology experiments.

Funding

This study was supported by the Korean Health Technology R&D Project, Ministry of Health & Welfare, Republic of Korea (HI13C1596) and a grant from the National Research Foundation (NRF-2016R1D1A3B03932491 and Stem Centric Co. Ltd., Republic of Korea.

Compliance with ethical standards

Competing interest

The authors declare that they have no competing interest.

Supplementary material

441_2019_3009_MOESM1_ESM.docx (13 kb)
ESM 1 (DOCX 13 kb)

References

  1. Andersson KE, Arner A (2004) Urinary bladder contraction and relaxation: physiology and pathophysiology. Physiol Rev 84:935–986CrossRefGoogle Scholar
  2. Bharti D, Shivakumar SB, Park JK, Ullah I, Subbarao RB, Park JS, Lee SL, Park BW, Rho GJ (2017) Comparative analysis of human Wharton’s jelly mesenchymal stem cells derived from different parts of the same umbilical cord. Cell Tissue Res 372:51–65CrossRefGoogle Scholar
  3. Corrao S, La Rocca G, Lo Iacono M, Corsello T, Farina F, Anzalone R (2013) Umbilical cord revisited: from Wharton’s jelly myofibroblasts to mesenchymal stem cells. Histol Histopathol 28:1235–1244Google Scholar
  4. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Dj P, Horwitz E (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. Int Soc Cell Ther Pos Statement Cytother 8:315–317Google Scholar
  5. Ghionzoli M, Repele A, Sartiani L, Costanzi G, Parenti A, Spinelli V, David AL, Garriboli M, Totonelli G, Tian J, Andreadis ST, Cerbai E, Mugelli A, Messineo A, Pierro A, Eaton S, De Coppi P (2013) Human amniotic fluid stem cell differentiation along smooth muscle lineage. FASEB J 27:4853–4865CrossRefGoogle Scholar
  6. Guo X, Stice SL, Boyd NL, Chen SY (2013) A novel in vitro model system for smooth muscle differentiation from human embryonic stem cell-derived mesenchymal cells. Am J Physiol Cell Physiol 304:C289–C298CrossRefGoogle Scholar
  7. Kobayashi K, Kubota T, Aso T (1998) Study on myofibroblast differentiation in the stromal cells of Wharton’s jelly: expression and localization of alpha-smooth muscle actin. Early Hum Dev 51:223–233CrossRefGoogle Scholar
  8. Moonen JR, Krenning G, Brinker MG, Koerts JA, van Luyn MJ, Harmsen MC (2010) Endothelial progenitor cells give rise to pro-angiogenic smooth muscle-like progeny. Cardiovasc Res 86:506–515CrossRefGoogle Scholar
  9. Ngo P, Ramalingam P, Phillips JA, Furuta GT (2006) Collagen gel contraction assay. Methods Mol Biol 341:103–109Google Scholar
  10. Rodríguez LV, Alfonso Z, Zhang R, Leung J, Wu B, Ignarro LJ (2006) Clonogenic multipotent stem cells in human adipose tissue differentiate into functional smooth muscle cells. Proc. Natl Acad Sci USA 103:12167–12172CrossRefGoogle Scholar
  11. Ross JJ, Hong Z, Willenbring B, Zeng L, Isenberg B, Eu HL, Reyes M, Keirstead SA, Weir EK, Tranquillo RT, Verfaillie CM (2006) Cytokine-induced differentiation of multipotent adult progenitor cells into functional smooth muscle cells. J Clin Invest 116:3139–3149CrossRefGoogle Scholar
  12. Shivakumar SB, Bharti D, Jang SJ, Hwang SC, Park JK, Shin JK, Byun JH, Park BW, Rho GJ (2015) Cryopreservation of human Wharton’s jelly-derived mesenchymal stem cells following controlled rate freezing protocol using different Cryoprotectants; a comparative study. Int J Stem Cells 8:155–169CrossRefGoogle Scholar
  13. Shivakumar SB, Bharti D, Subbarao RB, Jang SJ, Park JS, Ullah I, Park JK, Byun JH, Park BW, Rho GJ (2016) DMSO- and serum-free cryopreservation of Wharton’s jelly tissue isolated from human umbilical cord. J Cell Biochem 117:2397–2412CrossRefGoogle Scholar
  14. Song B, Jiang W, Alraies A, Liu Q, Gudla V, Oni J, Wei X, Sloan A, Ni L, Agarwal M (2016) Bladder smooth muscle cells differentiation from dental pulp stem cells: future potential for bladder tissue engineering. Stem Cells Int 2016:6979368.  https://doi.org/10.1155/2016/6979368 Google Scholar
  15. Subramanian A, Fong CY, Biswas A, Bongso A (2015) Comparative characterization of cells from the various compartments of the human umbilical cord shows that the Wharton’s jelly compartment provides the best source of clinically utilizable mesenchymal stem cells. PLoS One 10:e0127992CrossRefGoogle Scholar
  16. Takechi K, Kuwabara Y, Mizuno M (1993) Ultrastructural and immunohistochemical studies of Wharton’s jelly umbilical cord cells. Placenta 14:235–245CrossRefGoogle Scholar
  17. Wang C, Yin S, Cen L, Liu Q, Liu W, Cao Y, Cui L (2010) Differentiation of adipose-derived stem cells into contractile smooth muscle cells induced by transforming growth factor-beta1 and bone morphogenetic protein-4. Tissue Eng. Part A 16:1201–1213Google Scholar
  18. Wanjare M, Kuo F, Gerecht S (2013) Derivation and maturation of synthetic and contractile vascular smooth muscle cells from human pluripotent stem cells. Cardiovasc Res 97:321–330CrossRefGoogle Scholar
  19. Xu ZC, Zhang Q, Li H (2013) Human hair follicle stem cell differentiation into contractile smooth muscle cells is induced by transforming growth factor-β1 and platelet-derived growth factor BB. Mol Med Rep 8:1715–1721CrossRefGoogle Scholar
  20. Xu JG, Zhu SY, Heng BC, Dissanayaka WL, Zhang CF (2017) TGF-β1 induced differentiation of SHED into functional smooth muscle cells. Stem Cell Res Ther 8:1–10CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Dinesh Bharti
    • 1
  • Sharath Belame Shivakumar
    • 1
  • Young-Bum Son
    • 1
  • Young-Ho Choi
    • 1
  • Imran Ullah
    • 2
  • Hyeon-Jeong Lee
    • 1
  • Eun-Jin Kim
    • 3
  • Sun-A Ock
    • 2
  • Ji-Eun Park
    • 4
  • Ji-Kwon Park
    • 4
  • Dawon Kang
    • 3
  • Sung-Lim Lee
    • 1
  • Bong-Wook Park
    • 5
  • Gyu-Jin Rho
    • 1
    • 6
    Email author
  1. 1.Department of Theriogenology and Biotechnology, College of Veterinary MedicineGyeongsang National UniversityJinjuRepublic of Korea
  2. 2.Animal Biotechnology DivisionNational Institute of Animal Science, RDAWanju-gunRepublic of Korea
  3. 3.Department of Physiology and Institute of Health Sciences, School of MedicineGyeongsang National UniversityJinjuRepublic of Korea
  4. 4.Department of Obstetrics and Gynecology, School of MedicineGyeongsang National UniversityChangwonRepublic of Korea
  5. 5.Department of Oral and Maxillofacial Surgery, School of MedicineGyeongsang National UniversityChangwonRepublic of Korea
  6. 6.Research Institute of Life SciencesGyeongsang National UniversityJinjuRepublic of Korea

Personalised recommendations