Are umbilical cord mesenchymal stem cells advantageous over bone marrow mesenchymal stem cells for cell therapy in orthopaedics?

Mennan, C.; Owen, S.; Mayrogonatou, E.; Kletsas, D.; Oldershaw, R.; Richardson, J.; Roberts, S.

Abstract

Purpose: Mesenchymal Stem cells (MSC) from umbilical cords (UC) are of increasing interest for cell therapy in degenerative musculoskeletal disorders as they present little ethical consideration and are reported to contain immune privileged cells, which may be suitable for allogeneic based therapies. The use of MSCs as allogeneic cells in vivo would only be possible if they retained their immune privileged properties in an inflammatory environment. We have previously shown that cells obtained from UC are multipotent and have the recommended MSC cell surface marker profile according to the International Society for Cellular Therapy. The focus of this study was to characterise the immune properties of MSCs isolated from UC, particularly Wharton’s jelly (WJ) and cells from whole cord (MC) as we have previously shown that cells from these cord regions have the best multipotency. MC and WJ were compared to Bone Marrow MSCs (BMSCs) in order to utilise the most promising population of stem cells and to analyse the immune properties of these cells before and after stimulation with the pro-inflammatory cytokine, interferon-γ (IFN-γ).

Methods: Umbilical cords were dissected to obtain WJ and MC was obtained from a ∼2 cm section of whole cord, which was minced into small (∼2mm2) pieces and digested in collagenase I for 1 h at 37°C. Resulting cells were cultured in DMEM-F12, 10 % FCS and Penicillin/streptomycin. Flow cytometry, immunocytochemistry and Western blotting was used to characterise cell markers indicative of pluripotency (SOX2, Nanog, REX-1, OCT3/4, SSEA-3, 4, TRA-1-60, TRA-1-81 and alkaline phosphatase) and immunogenicity (using co-stimulatory markers, CD40, 80, 86, in addition to HLA-G, Indoleamine 2 3-dioxygenase and HLA-DR) before and after stimulation with IFN-γ (500 ng/ml for 48 h). BMSC and UCMSC were cultured with allogeneic CD4+ T cells (responder cells) (at a ratio of 1:5) labelled with violet proliferation dye in addition to peripheral blood mononuclear cells (PBMC) (used as stimulator cells) for 5 days. Controls were CD4+ T cells and PBMCs alone. After 5 days T cells were analysed via flow cytometry to assess their proliferative response.

Results: All cells showed positivity for the markers SOX2, Nanog, REX-1, SSEA-4, Alkaline phosphatase, TRA-1-81, HLA-G and were negative for OCT 3/4, SSEA-3 and the co-stimulatory markers CD40, CD80, CD86. BMSC were positive for TRA-1-60, whereas UC MSC were negative. Differences were seen between BMSC and UC MSC after stimulation with IFN-γ. UC MSC remained negative for the co-stimulatory markers CD40, 80, 86 and HLA-DR after IFN-γ stimulation, but, BMSC showed up-regulation of HLA-DR after exposure to IFN-γ. All cells were negative for IDO before treatment with IFN-γ and became positive after stimulation. T cell proliferation was found to be supressed in all cultures containing either BMSCs or UC MSCs, showing that cells from both of these sources have immunomodulatory properties and may be capable of dampening down immune response in vivo.

Conclusions: Umbilical cord derived MSCs may have more promise than BMSCs as an allogeneic cell therapy as they do not produce co-stimulatory markers or MHC (major histocompatibility complex) class II antigens after culture in vitro in an inflammatory environment such as that which may be found in an osteoarthritic joint.

Citation

Mennan, C., Owen, S., Mayrogonatou, E., Kletsas, D., Oldershaw, R., Richardson, J., & Roberts, S. (2014). Are umbilical cord mesenchymal stem cells advantageous over bone marrow mesenchymal stem cells for cell therapy in orthopaedics?. . https://doi.org/10.1016/j.joca.2014.02.924

• Conference Name: 2014 World Congress on Osteoarthritis: Promoting Clinical and Basic Research in Osteoarthritis
• Conference Location: Paris, France
• Start Date: Apr 24, 2014
• End Date: Apr 27, 2014
• Online Publication Date: Mar 20, 2014
• Publication Date: 2014-04
• Deposit Date: Jun 8, 2023
• Publisher: Elsevier
• Volume: 22
• Pages: S486-S487
• DOI: https://doi.org/10.1016/j.joca.2014.02.924
• Keywords: Orthopedics and Sports Medicine; Biomedical Engineering; Rheumatology