Wharton’s Jelly Derived Mesenchymal Stem Cells: Comparing Human and Horse

  • Barbara Merlo
  • Gabriella Teti
  • Eleonora Mazzotti
  • Laura Ingrà
  • Viviana Salvatore
  • Marina Buzzi
  • Giorgia Cerqueni
  • Manuela Dicarlo
  • Aliai Lanci
  • Carolina Castagnetti
  • Eleonora Iacono
Article

Abstract

Wharton’s jelly (WJ) is an important source of mesenchymal stem cells (MSCs) both in human and other animals. The aim of this study was to compare human and equine WJMSCs. Human and equine WJMSCs were isolated and cultured using the same protocols and culture media. Cells were characterized by analysing morphology, growth rate, migration and adhesion capability, immunophenotype, differentiation potential and ultrastructure. Results showed that human and equine WJMSCs have similar ultrastructural details connected with intense synthetic and metabolic activity, but differ in growth, migration, adhesion capability and differentiation potential. In fact, at the scratch assay and transwell migration assay, the migration ability of human WJMSCs was higher (P < 0.05) than that of equine cells, while the volume of spheroids obtained after 48 h of culture in hanging drop was larger than the volume of equine ones (P < 0.05), demonstrating a lower cell adhesion ability. This can also revealed in the lower doubling time of equine cells (3.5 ± 2.4 days) as compared to human (6.5 ± 4.3 days) (P < 0.05), and subsequently in the higher number of cell doubling after 44 days of culture observed for the equine (20.3 ± 1.7) as compared to human cells (8.7 ± 2.4) (P < 0.05), and to the higher (P < 0.05) ability to form fibroblast colonies at P3. Even if in both species tri-lineage differentiation was achieved, equine cells showed an higher chondrogenic and osteogenic differentiation ability (P < 0.05). Our findings indicate that, although the ultrastructure demonstrated a staminal phenotype in human and equine WJMSCs, they showed different properties reflecting the different sources of MSCs.

Keywords

Mesenchymal stem cell Wharton’s jelly Horse Human Transmission electron microscopy 

Notes

Acknowledgements

The author wish to thank Mirella Falconi, manager of “Laboratorio di morfologia e biologia cellulare e tissutale” (Department for Biomedical and Neuromotor Sciences). This research was funded by RFO (Ricerca Fondamentale Orientata), University of Bologna.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Dominici, M., Le Blanc, K., Mueller, I., et al. (2006). Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8, 315–317.CrossRefPubMedGoogle Scholar
  2. 2.
    Vangsness, C. T. Jr., Sternberg, H., & Harris, L. (2015). Umbilical cord tissue offers the greatest number of harvestable mesenchymal stem cells for research and clinical application: a literature review of different harvest sites. Arthroscopy, 31, 1836–1843.CrossRefPubMedGoogle Scholar
  3. 3.
    Iacono, E., Rossi, B., & Merlo, B. (2015). Stem cells from foetal adnexa and fluid in domestic animals: an update on their features and clinical application. Reproduction in Domestic Animals, 50, 353–364.CrossRefPubMedGoogle Scholar
  4. 4.
    Troyer, D. L., & Weiss, M. L. (2008). Wharton’s jelly-derived cells are a primitive stromal cell population. Stem Cells, 26, 591–599.CrossRefPubMedGoogle Scholar
  5. 5.
    Harding, J., Roberts, R. M., & Mirochnitchenko, O. (2013). Large animal models for stem cell therapy. Stem Cell Research & Therapy, 4, 23.CrossRefGoogle Scholar
  6. 6.
    Rainaldi, G., Pinto, B., Piras, A., Vatteroni, L., Simi, S., & Citti, L. (1991). Reduction of proliferative heterogeneity of CHEF18 Chinese hamster cell line during the progression toward tumorigenicity. In Vitro Cellular & Development Biology, 27, 949–952.CrossRefGoogle Scholar
  7. 7.
    Piccinini, F., Tesei, A., Arienti, C., & Bevilacqua, A. (2015). Cancer multicellular spheroids: volume assessment from a single 2D projection. Computer Methods and Programs in Biomedicine, 118, 95–106.CrossRefPubMedGoogle Scholar
  8. 8.
    De Schauwer, C., Piepers, S., Van de Walle, G. R., et al. (2012). In search for cross-reactivity to immunophenotype equine mesenchymal stromal cells by multicolor flow cytometry. Cytometry Part A, 81, 312–323.CrossRefGoogle Scholar
  9. 9.
    Ibrahim, S., & Steinbach, F. (2012). Immunoprecipitation of equine CD molecules using anti-human MABs previously analyzed by flow cytometry and immunohistochemistry. Veterinary Immunology and Immunopathology, 145, 7–13.CrossRefPubMedGoogle Scholar
  10. 10.
    Iacono, E., Merlo, B., Romagnoli, N., Rossi, B., Ricci, F., & Spadari, A. (2015). Equine bone marrow and adipose tissue mesenchymal stem cells: cytofluorimetric characterization, in vitro differentiation, and clinical application. Journal of Equine Veterinary Science, 35, 130–140.CrossRefGoogle Scholar
  11. 11.
    Patterson-Kane, J. C., Becker, D. L., & Rich, T. (2012). The pathogenesis of tendon microdamage in athletes: the horse as a natural model for basic cellular research. Journal of Comparative Pathology, 147, 227–247.CrossRefPubMedGoogle Scholar
  12. 12.
    Smith, R. K. W. (2008). Mesenchymal stem cell therapy for equine tendinopathy. Disability and Rehabilitation, 30, 1752–1758.CrossRefPubMedGoogle Scholar
  13. 13.
    Crovace, A., Lacitignola, L., Rossi, G., & Francioso, E. (2010). Histological and immunohistochemical evaluation of autologous cultured bone marrow mesenchymal stem cells and bone marrow mononucleated cells in collagenase-induced tendinitis of equine superficial digital flexor tendon. Veterinary Medicine International, 250978.Google Scholar
  14. 14.
    Godwin, E. E., Young, N. J., Dudhia, J., Beamish, I. C., & Smith, R. K. (2012). Implantation of bone marrow-derived mesenchymal stem cells demonstrates improved outcome in horses with overstrain injury of the superficial digital flexor tendon. Equine Veterinary Journal, 44, 25–32.CrossRefPubMedGoogle Scholar
  15. 15.
    Pasquinelli, G., Tazzari, P., Ricci, F., et al. (2007). Ultrastructural characteristics of human mesenchymal stromal (stem) cells derived from bone marrow and term placenta. Ultrastructural Pathology, 31, 23–31.CrossRefPubMedGoogle Scholar
  16. 16.
    Orsolic, I., Jurada, D., Pullen, N., Oren, M., Eliopoulos, A. G., & Volarevic, S. (2016). The relationship between the nucleolus and cancer: current evidence and emerging paradigms. Seminars in Cancer Biology, 37–38, 36–50.CrossRefPubMedGoogle Scholar
  17. 17.
    Pascucci, L., Mercati, F., Marini, C., et al. (2010). Ultrastructural morphology of equine adipose-derived mesenchymal stem cells. Histology and Histopathology, 25, 1277–1285.PubMedGoogle Scholar
  18. 18.
    Teti, G., Cavallo, C., Grigolo, B., et al. (2012). Ultrastructural analysis of human bone marrow mesenchymal stem cells during in vitro osteogenesis and chondrogenesis. Microscopy Research and Technique, 75, 596–604.CrossRefPubMedGoogle Scholar
  19. 19.
    García-Prat, L., Martínez-Vicente, M., & Muñoz-Cánoves, P. (2016). Autophagy: a decisive process for stemness. Oncotarget, 7, 12286–12288.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Sbrana, F. V., Cortini, M., Avnet, S., et al. (2016). The role of autophagy in the maintenance of stemness and differentiation of mesenchymal stem cells. Stem Cell Reviews, 12, 621–633.CrossRefPubMedGoogle Scholar
  21. 21.
    Iacono, E., Brunori, L., Pirrone, A., et al. (2012). Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton’s jelly in the horse. Reproduction, 143, 455–468.CrossRefPubMedGoogle Scholar
  22. 22.
    Mattioli-Belmonte, M., Teti, G., Salvatore, V., et al. (2015). Stem cell origin differently affects bone tissue engineering strategies. Frontiers in Physiology, 6, 266.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Li, G., Zhang, X. A., Wang, H., et al. (2009). Comparative proteomic analysis of mesenchymal stem cells derived from human bone marrow, umbilical cord, and placenta: Implication in the migration. Proteomics, 9, 20–30.CrossRefPubMedGoogle Scholar
  24. 24.
    Sart, S., Tsai, A. C., Li, Y., & Ma, T. (2014). Three-dimensional aggregates of mesenchymal stem cells: cellular mechanism, biological properties, and applications. Tissue Engineering Part B Reviews, 20, 365–380.CrossRefPubMedGoogle Scholar
  25. 25.
    Wang, W., Itaka, K., Ohba, S., et al. (2009). 3D spheroid culture system on micropatterned substrates for improved differentiation efficiency of multipotent mesenchymal stem cells. Biomaterials, 30, 2705–2715.CrossRefPubMedGoogle Scholar
  26. 26.
    Suzuki, S., Muneta, T., Tsuji, K., et al. (2012). Properties and usefulness of aggregates of synovial mesenchymal stem cells as a source for cartilage regeneration. Arthritis Research & Therapy, 14, R136.CrossRefGoogle Scholar
  27. 27.
    Mizuno, H., & Hyakusoku, H. (2003). Mesengenic potential and future clinical perspective of human processed lipospirated cells. Journal of Nippon Medical School, 70, 300–306.CrossRefPubMedGoogle Scholar
  28. 28.
    Janderova, L., McNeil, M., Murrell, A. N., Mynatt, R. L., & Smith, S. R. (2003). Human mesenchymal stem cells as an in vitro model for human adipogenesis. Obesity Research, 11, 65–74.CrossRefPubMedGoogle Scholar
  29. 29.
    De Schauwer, C., Meyer, E., Van de Walle, G. R., & Van Soom, A. (2011). Markers of stemness in equine mesenchymal stem cells: a plea for uniformity. Theriogenology, 75, 1431–1443.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Barbara Merlo
    • 1
  • Gabriella Teti
    • 2
  • Eleonora Mazzotti
    • 3
  • Laura Ingrà
    • 2
  • Viviana Salvatore
    • 4
  • Marina Buzzi
    • 5
  • Giorgia Cerqueni
    • 6
  • Manuela Dicarlo
    • 6
  • Aliai Lanci
    • 1
  • Carolina Castagnetti
    • 1
  • Eleonora Iacono
    • 1
  1. 1.Department of Veterinary Medical SciencesUniversity of BolognaOzzano EmiliaItaly
  2. 2.Department for Biomedical and Neuromotor SciencesUniversity of BolognaBolognaItaly
  3. 3.Department of Comparative Biomedical SciencesUniversity of TeramoTeramoItaly
  4. 4.An2H Discovery Limited, National Institute for Cellular Bioetchnology (NICB)Dublin City University CampusGlasnevinIreland
  5. 5.Banca dei Tessuti, del Sangue cordonale e Biobanca Policlinico S.Orsola-MalpighiBolognaItaly
  6. 6.Department of Clinical and Molecular SciencesPolytechnic University of MarcheAnconaItaly

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