pp 1-13 | Cite as

Isolation, Expansion, and Characterization of Wharton’s Jelly-Derived Mesenchymal Stromal Cell: Method to Identify Functional Passages for Experiments

  • Shuh-Wen Aung
  • Noor Hayaty Abu Kasim
  • Thamil Selvee RamasamyEmail author
Part of the Methods in Molecular Biology book series


The therapeutic potential of human mesenchymal stromal stem cells (hMSCs) for cell-based therapeutic is greatly influenced by the in vitro culture condition including the culture conditions. Nevertheless, there are many technical challenges needed to be overcome prior to the clinical use including the quantity, quality, and heterogeneity of the cells. Therefore, it is necessary to develop a stem cell culture procedure or protocol for cell expansion in order to generate reproducible and high-quality cells in accordance with good manufacturing practice for clinical and therapeutic purposes. Here we assessed the MSCs characteristic of human Wharton’s jelly mesenchymal stromal cells in in vitro culture according to the criteria established by the International Society for Cellular Therapy. Besides, the viability of the WJMSCs was determined in order to increase the confidence that the cells are employed to meet the therapeutic efficacy.


Wharton’s jelly Mesenchymal stromal cells In vitro passaging Replication senescence Stemness 



This research was supported by High Impact Research MOHE Grant UM.C/625/1/HIR/MOHE/DENT/01 from Ministry of Higher Education Malaysia, Fundamental Research Grant Scheme (FRGS FP044-2014B) from Ministry of Education, Malaysia, and University of Malaya Research Grant (RP019C-13HTM) from University of Malaya.


  1. 1.
    Heathman TR, Nienow AW, McCall MJ, Coopman K, Kara B, Hewitt CJ (2015) The translation of cell-based therapies: clinical landscape and manufacturing challenges. Regen Med 10:49–64Google Scholar
  2. 2.
    Buzhor E, Leshansky L, Blumenthal J, Barash H, Warshawsky D, Mazor Y, Shtrichman R (2014) Cell-based therapy approaches: the hope for incurable diseases. Regen Med 9:649–672Google Scholar
  3. 3.
    Wei X, Yang X, Han ZP, Qu FF, Shao L, Shi YF (2013) Mesenchymal stem cells: a new trend for cell therapy. Acta Pharmacol Sin 34:747–754Google Scholar
  4. 4.
    Kalaszczynska I, Ferdyn K (2015) Wharton’s jelly derived mesenchymal stem cells: future of regenerative medicine? Recent findings and clinical significance. Biomed Res Int 2015:430847Google Scholar
  5. 5.
    Batsali AK, Kastrinaki MC, Papadaki HA, Pontikoglou C (2013) Mesenchymal stem cells derived from Wharton’s Jelly of the umbilical cord: biological properties and emerging clinical applications. Curr Stem Cell Res Ther 8:144–155Google Scholar
  6. 6.
    Galipeau J, Sensebe L (2018) Mesenchymal stromal cells: clinical challenges and therapeutic opportunities. Cell Stem Cell 22:824–833Google Scholar
  7. 7.
    Trounson A, McDonald C (2015) Stem cell therapies in clinical trials: progress and challenges. Cell Stem Cell 17:11–22Google Scholar
  8. 8.
    Sepulveda JC, Tome M, Fernandez ME, Delgado M, Campisi J, Bernad A, Gonzalez MA (2014) Cell senescence abrogates the therapeutic potential of human mesenchymal stem cells in the lethal endotoxemia model. Stem Cells 32:1865–1877Google Scholar
  9. 9.
    Wagner W, Horn P, Castoldi M, Diehlmann A, Bork S, Saffrich R, Benes V, Blake J, Pfister S, Eckstein V et al (2008) Replicative senescence of mesenchymal stem cells: a continuous and organized process. PLoS One 3:e2213Google Scholar
  10. 10.
    Wagner W, Ho AD, Zenke M (2010) Different facets of aging in human mesenchymal stem cells. Tissue Eng Part B Rev 16:445–453Google Scholar
  11. 11.
    Wagner W, Bork S, Lepperdinger G, Joussen S, Ma N, Strunk D, Koch C (2010) How to track cellular aging of mesenchymal stromal cells? Aging 2:224–230Google Scholar
  12. 12.
    Hayflick L (1965) The limited in vitro lifetime of human diploid cell strains. Exp Cell Res 37:614–636Google Scholar
  13. 13.
    Wagner W, Bork S, Horn P, Krunic D, Walenda T, Diehlmann A, Benes V, Blake J, Huber FX, Eckstein V et al (2009) Aging and replicative senescence have related effects on human stem and progenitor cells. PLoS One 4:e5846Google Scholar
  14. 14.
    Turinetto V, Vitale E, Giachino C (2016) Senescence in human mesenchymal stem cells: functional changes and implications in stem cell-based therapy. Int J Mol Sci 17(7)Google Scholar
  15. 15.
    Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop D, Horwitz E (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8:315–317Google Scholar
  16. 16.
    Kurz DJ, Decary S, Hong Y, Erusalimsky JD (2000) Senescence-associated (beta)-galactosidase reflects an increase in lysosomal mass during replicative ageing of human endothelial cells. J Cell Sci 113:3613–3622Google Scholar

Copyright information

© Springer Science+Business Media New York 2019

Authors and Affiliations

  • Shuh-Wen Aung
    • 1
    • 2
  • Noor Hayaty Abu Kasim
    • 1
    • 2
  • Thamil Selvee Ramasamy
    • 3
    Email author
  1. 1.Department of Restorative Dentistry, Faculty of DentistryUniversity of MalayaKuala LumpurMalaysia
  2. 2.Regenerative Dentistry Research Group, Faculty of DentistryUniversity of MalayaKuala LumpurMalaysia
  3. 3.Stem Cell Biological Laboratory, Department of Molecular Medicine, Faculty of MedicineUniversity of MalayaKuala LumpurMalaysia

Personalised recommendations