Skip to main content

Advertisement

Log in

Phenotypic and Functional Characterization of Mesenchymal Stem Cells from Chorionic Villi of Human Term Placenta

  • Published:
Stem Cell Reviews and Reports Aims and scope Submit manuscript

Abstract

Background

Bone marrow derived mesenchymal stem cells (BM-MSCs) are used extensively in transplantation but their use is associated with many problems including low abundance in BM, low overall number, decreased differentiation potential with age and the invasive isolation procedures needed to obtain BM. We report a novel method of isolating placental MSCs (pMSCs) from chorionic villi, which exhibit the phenotypic and functional characteristics that will make them an attractive source of MSCs for cell-based therapy.

Methods

A novel explant approach was used to isolate pMSCs from chorionic villi of human placentae. These pMSCs were characterized by flow cytometry and were differentiated into adipocytes, osteocytes and chondrocytes using differentiation medium as demonstrated by cytochemical staining. The gene and protein expression profiles of pMSCs were also characterized using real time polymerase chain reaction (PCR) and flow cytometry, respectively. In addition, cytokine secretion by pMSCs was also analysed using sandwich enzyme-linked immunosorbent assay (ELISA) technique. Moreover, the migration and proliferation potentials of pMSCs were also determined.

Results

pMSCs were isolated from fetal part of the chorionic villi and these pMSCs expressed CD44, CD90, CD105, CD146, CD166 and HLA-ABC but not CD14, CD19, CD40, CD45, CD80, CD83, CD86 and HLA-DR. In addition, these pMSCs differentiated into osteocytes, chondrocytes and adipocytes and they also expressed several adhesion molecules, chemokines/receptors, growth factor receptors and cytokines/receptors. Moreover, they secreted many cytokines (IL-1Ra, IL6, IL8, IL10, IL11 and IL15) and they were able to proliferate. Furthermore, they migrated in response to chemotactic factors including stromal cell-derived factor-1 (SDF-1), platelet derived growth factor (PDGF), hepatocyte growth factor (HGF), and monocyte chemotactic protein-1 (MCP-1).

Conclusions

We devised a novel explant method of isolating pMSCs that expressed many biological factors responsible for mediating cellular processes such as migration/homing, immune modulation and angiogenesis. Therefore, we suggest that pMSCs prepared from human term placental chorionic villous explants are an attractive source of MSCs for cell therapy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Friedenstein, A. J., Chailakhjan, R. K., & Lalykina, K. S. (1970). The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells. Cell and Tissue Kinetics, 3, 393–403.

    CAS  PubMed  Google Scholar 

  2. Augello, A., Kurth, T. B., & De Bari, C. (2010). Mesenchymal stem cells: a perspective from in vitro cultures to in vivo migration and niches. European Cells & Materials, 20, 121–133.

    CAS  Google Scholar 

  3. Krampera, M., Pizzolo, G., Aprili, G., & Franchini, M. (2006). Mesenchymal stem cells for bone, cartilage, tendon and skeletal muscle repair. Bone, 39, 678–683.

    CAS  PubMed  Google Scholar 

  4. Phinney, D. G., & Prockop, D. J. (2007). Concise review: mesenchymal stem/multipotent stromal cells: the state of transdifferentiation and modes of tissue repair—current views. Stem Cells, 25, 2896–2902.

    PubMed  Google Scholar 

  5. Tanaka, M., & Miyajima, A. (2012). Identification and isolation of adult liver stem/progenitor cells. Methods in Molecular Biology, 826, 25–32.

    CAS  PubMed  Google Scholar 

  6. Sakaguchi, Y., Sekiya, I., Yagishita, K., & Muneta, T. (2005). Comparison of human stem cells derived from various mesenchymal tissues: superiority of synovium as a cell source. Arthritis and Rheumatism, 52, 2521–2529.

    PubMed  Google Scholar 

  7. Roubelakis, M. G., Pappa, K. I., Bitsika, V., et al. (2007). Molecular and proteomic characterization of human mesenchymal stem cells derived from amniotic fluid: comparison to bone marrow mesenchymal stem cells. Stem Cells and Development, 16, 931–952.

    CAS  PubMed  Google Scholar 

  8. In’t Anker, P. S., Scherjon, S. A., Kleijburg-van der Keur, C., et al. (2004). Isolation of mesenchymal stem cells of fetal or maternal origin from human placenta. Stem Cells, 22, 1338–1345.

    Google Scholar 

  9. Kanematsu, D., Shofuda, T., Yamamoto, A., et al. (2011). Isolation and cellular properties of mesenchymal cells derived from the decidua of human term placenta. Differentiation; Research in Biological Diversity, 82, 77–88.

    CAS  PubMed  Google Scholar 

  10. Pelagiadis, I., Relakis, K., Kalmanti, L., & Dimitriou, H. (2012). CD133 immunomagnetic separation: effectiveness of the method for CD133(+) isolation from umbilical cord blood. Cytotherapy.

  11. Gronthos, S., Arthur, A., Bartold, P. M., & Shi, S. (2011). A method to isolate and culture expand human dental pulp stem cells. Methods in Molecular Biology, 698, 107–121.

    CAS  PubMed  Google Scholar 

  12. 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.

    CAS  PubMed  Google Scholar 

  13. Le Blanc, K., Frassoni, F., Ball, L., et al. (2008). Mesenchymal stem cells for treatment of steroid-resistant, severe, acute graft-versus-host disease: a phase II study. Lancet, 371, 1579–1586.

    PubMed  Google Scholar 

  14. Toubai, T., Paczesny, S., Shono, Y., et al. (2009). Mesenchymal stem cells for treatment and prevention of graft-versus-host disease after allogeneic hematopoietic cell transplantation. Current Stem Cell Research & Therapy, 4, 252–259.

    CAS  Google Scholar 

  15. Majumdar, M. K., Thiede, M. A., Haynesworth, S. E., Bruder, S. P., & Gerson, S. L. (2000). Human marrow-derived mesenchymal stem cells (MSCs) express hematopoietic cytokines and support long-term hematopoiesis when differentiated toward stromal and osteogenic lineages. Journal of Hematotherapy & Stem Cell Research, 9, 841–848.

    CAS  Google Scholar 

  16. D’Ippolito, G., Schiller, P. C., Ricordi, C., Roos, B. A., & Howard, G. A. (1999). Age-related osteogenic potential of mesenchymal stromal stem cells from human vertebral bone marrow. Journal of Bone and Mineral Research: The Official Journal of the American Society for Bone and Mineral Research, 14, 1115–1122.

    Google Scholar 

  17. Brooke, G., Tong, H., Levesque, J. P., & Atkinson, K. (2008). Molecular trafficking mechanisms of multipotent mesenchymal stem cells derived from human bone marrow and placenta. Stem Cells and Development, 17, 929–940.

    CAS  PubMed  Google Scholar 

  18. Fukuchi, Y., Nakajima, H., Sugiyama, D., Hirose, I., Kitamura, T., & Tsuji, K. (2004). Human placenta-derived cells have mesenchymal stem/progenitor cell potential. Stem Cells, 22, 649–658.

    CAS  PubMed  Google Scholar 

  19. Yen, B. L., Huang, H. I., Chien, C. C., et al. (2005). Isolation of multipotent cells from human term placenta. Stem Cells, 23, 3–9.

    CAS  PubMed  Google Scholar 

  20. Ringe, J., Strassburg, S., Neumann, K., et al. (2007). Towards in situ tissue repair: human mesenchymal stem cells express chemokine receptors CXCR1, CXCR2 and CCR2, and migrate upon stimulation with CXCL8 but not CCL2. Journal of Cellular Biochemistry, 101, 135–146.

    CAS  PubMed  Google Scholar 

  21. Li, D., Wang, G. Y., Dong, B. H., Zhang, Y. C., Wang, Y. X., & Sun, B. C. (2007). Biological characteristics of human placental mesenchymal stem cells and their proliferative response to various cytokines. Cells, Tissues, Organs, 186, 169–179.

    CAS  PubMed  Google Scholar 

  22. Ponte, A. L., Marais, E., Gallay, N., et al. (2007). The in vitro migration capacity of human bone marrow mesenchymal stem cells: comparison of chemokine and growth factor chemotactic activities. Stem Cells, 25, 1737–1745.

    CAS  PubMed  Google Scholar 

  23. Zhang, X., Mitsuru, A., Igura, K., et al. (2006). Mesenchymal progenitor cells derived from chorionic villi of human placenta for cartilage tissue engineering. Biochemical and Biophysical Research Communications, 340, 944–952.

    CAS  PubMed  Google Scholar 

  24. Igura, K., Zhang, X., Takahashi, K., Mitsuru, A., Yamaguchi, S., & Takashi, T. A. (2004). Isolation and characterization of mesenchymal progenitor cells from chorionic villi of human placenta. Cytotherapy, 6, 543–553.

    CAS  PubMed  Google Scholar 

  25. Hirota, M., Ohmuraya, M., & Baba, H. (2006). The role of trypsin, trypsin inhibitor, and trypsin receptor in the onset and aggravation of pancreatitis. Journal of Gastroenterology, 41, 832–836.

    CAS  PubMed  Google Scholar 

  26. Castrechini, N. M., Murthi, P., Gude, N. M., et al. (2010). Mesenchymal stem cells in human placental chorionic villi reside in a vascular Niche. Placenta, 31, 203–212.

    CAS  PubMed  Google Scholar 

  27. Gronthos, S., Franklin, D. M., Leddy, H. A., Robey, P. G., Storms, R. W., & Gimble, J. M. (2001). Surface protein characterization of human adipose tissue-derived stromal cells. Journal of Cellular Physiology, 189, 54–63.

    CAS  PubMed  Google Scholar 

  28. Ji, J. F., He, B. P., Dheen, S. T., & Tay, S. S. (2004). Interactions of chemokines and chemokine receptors mediate the migration of mesenchymal stem cells to the impaired site in the brain after hypoglossal nerve injury. Stem Cells, 22, 415–427.

    CAS  PubMed  Google Scholar 

  29. Murphy, P. M., Baggiolini, M., Charo, I. F., et al. (2000). International union of pharmacology. XXII. Nomenclature for chemokine receptors. Pharmacological Reviews, 52, 145–176.

    CAS  PubMed  Google Scholar 

  30. Kim, D. H., Yoo, K. H., Choi, K. S., et al. (2005). Gene expression profile of cytokine and growth factor during differentiation of bone marrow-derived mesenchymal stem cell. Cytokine, 31, 119–126.

    CAS  PubMed  Google Scholar 

  31. Segers, V. F., Van Riet, I., Andries, L. J., et al. (2006). Mesenchymal stem cell adhesion to cardiac microvascular endothelium: activators and mechanisms. American Journal of Physiology. Heart and Circulatory Physiology, 290, H1370–H1377.

    CAS  PubMed  Google Scholar 

  32. Yong, K. L., Watts, M., Shaun Thomas, N., Sullivan, A., Ings, S., & Linch, D. C. (1998). Transmigration of CD34+ cells across specialized and nonspecialized endothelium requires prior activation by growth factors and is mediated by PECAM-1 (CD31). Blood, 91, 1196–1205.

    CAS  PubMed  Google Scholar 

  33. Kagimoto, Y., Yamada, H., Ishikawa, T., et al. (2008). A regulatory role of interleukin 15 in wound healing and mucosal infection in mice. Journal of Leukocyte Biology, 83, 165–172.

    CAS  PubMed  Google Scholar 

  34. Abumaree, M., Al Jumah, M., Pace, R. A., & Kalionis, B. (2011). Immunosuppressive properties of mesenchymal stem cells. Stem Cell Reviews.

  35. Perrigoue, J. G., Zaph, C., Guild, K., Du, Y., & Artis, D. (2009). IL-31-IL-31R interactions limit the magnitude of Th2 cytokine-dependent immunity and inflammation following intestinal helminth infection. Journal of Immunology, 182, 6088–6094.

    CAS  Google Scholar 

  36. Artis, D., Villarino, A., Silverman, M., et al. (2004). The IL-27 receptor (WSX-1) is an inhibitor of innate and adaptive elements of type 2 immunity. Journal of Immunology, 173, 5626–5634.

    CAS  Google Scholar 

  37. Nold, M. F., Nold-Petry, C. A., Pott, G. B., et al. (2008). Endogenous IL-32 controls cytokine and HIV-1 production. Journal of Immunology, 181, 557–565.

    CAS  Google Scholar 

  38. Andoh, A., Shioya, M., Nishida, A., et al. (2009). Expression of IL-24, an activator of the JAK1/STAT3/SOCS3 cascade, is enhanced in inflammatory bowel disease. Journal of Immunology, 183, 687–695.

    CAS  Google Scholar 

  39. Su, Z., Emdad, L., Sauane, M., et al. (2005). Unique aspects of mda-7/IL-24 antitumor bystander activity: establishing a role for secretion of MDA-7/IL-24 protein by normal cells. Oncogene, 24, 7552–7566.

    CAS  PubMed  Google Scholar 

  40. Suh, W. K., Gajewska, B. U., Okada, H., et al. (2003). The B7 family member B7-H3 preferentially down-regulates T helper type 1-mediated immune responses. Nature Immunology, 4, 899–906.

    CAS  PubMed  Google Scholar 

  41. Starke, A., Lindenmeyer, M. T., Segerer, S., et al. (2010). Renal tubular PD-L1 (CD274) suppresses alloreactive human T-cell responses. Kidney International, 78, 38–47.

    CAS  PubMed  Google Scholar 

  42. Singh, A. K., Stock, P., & Akbari, O. (2011). Role of PD-L1 and PD-L2 in allergic diseases and asthma. Allergy, 66, 155–162.

    CAS  PubMed  Google Scholar 

  43. Jones, B. J., Brooke, G., Atkinson, K., & McTaggart, S. J. (2007). Immunosuppression by placental indoleamine 2,3-dioxygenase: a role for mesenchymal stem cells. Placenta, 28, 1174–1181.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We would also like to thank the staff and patients of the Delivery Unit, King Abdul Aziz Medical City for their help in obtaining placentae. This study was supported by grants from King Abdulla International Medical Research Centre (Grant No. RC08/114) and King Abdulaziz City for Science and Technology (Grant No. ARP-29-186). B. Kalionis acknowledges that support of National Health and Medical Research Grant 509178.

Conflict of interest The authors declare no potential conflicts of interest.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to M. H. Abumaree.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Abumaree, M.H., Al Jumah, M.A., Kalionis, B. et al. Phenotypic and Functional Characterization of Mesenchymal Stem Cells from Chorionic Villi of Human Term Placenta. Stem Cell Rev and Rep 9, 16–31 (2013). https://doi.org/10.1007/s12015-012-9385-4

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12015-012-9385-4

Keywords

Navigation