Umbilical cord mesenchymal stem cell transplantation in the treatment of multiple sclerosis

Abstract

To investigate the clinical efficacy and safety of umbilical cord mesenchymal stem cell (UCMSC) transplantation for treating multiple sclerosis (MS), the patients with MS were recruited and treated with UCMSC. The procedure of preparing UCMSC was in accordance with the standards formulated by the International Society for Cell Biology. Cell surface markers, multiple differentiation potential and safety of UCMSC for transplantation were detected. The number of cells in each infusion was 1 to 2×10⁶ cells/kg. Patients were recruited in accordance with the standards of the International Mesenchymal Stem Cells Transplantation Study Group. After treatment, the clinical therapeutic effects including symptoms, vital signs, clinical attacks, magnetic resonance imaging (MRI), neurological function scores and adverse reactions such as fever, dizziness, and vascular irritation were monitored and evaluated. In addition, the regulatory effects of UCMSC on immune system of patients were also assessed. The results showed that the patients' symptoms were improved after UCMSC transplantation. No clinical attacks occurred during transplantation. MRI revealed a reduced number of foci and Expanded Disability Status Scale scores were decreased. Some of patients had adverse reactions after transplantation. These adverse effects were not serious and lasted short duration, thus no intervention was conducted and let it be eliminated by itself. The mRNA expression of CD86, IL-2, CTLA-4, and HLADRB1 in peripheral blood was significantly decreased after UCMSC transplantation (P < 0.05). Based on our present studies, UCMSCs would be considered as a safe and alternative option for treatment of MS.

Keywords: Umbilical cord mesenchymal stem cell; clinical efficacy; multiple sclerosis; transplantation.

Figure 1

Multi-differentiation of UCMSCs in vitro. A. Undifferentiated UCMSCs displayed fibroblast-like cell morphology. B. Von Kossa staining for osteogenic differentiation. C. Oil red O staining for adipogenic differentiation. D. Neurofilament M immunofluorescence staining for neurogenic differentiation. E-P. Immunophenotypic characterization of UCMSCs by flow cytometry. The cells were positive for CD49, CD90, CD29, CD271, CD73, CD105, CD166, and CD44 but negative for CD34, CD45, CD123, and HLA-DR.

Figure 2

A, B. UCMSCs from passages 7 and 23 presented a normal chromosome karyotype with the absence of polyploidy. C. The representative photo of mice inoculated with UCMSC after 8 weeks, no tumor formation. D. The representative photo of mice inoculated with SPC-A-1 as the positive control.

Figure 3

Evaluation of changes in EDSS scores in three multiple sclerosis patients at different time points. In Patient 2, the symptoms were stable, and the EDSS score was 6.5. The EDSS score of patient in control group was 7.0, after received drug treatment, indicating the disease was continuously progressive.

Figure 4

MRI images of patients in the cerebral transverse plane and sagittal cervical segment at different time points. A. Patient 1, after patient 1 received seven times of treatments, lesions on the right side of the cerebral ventricle became pale and tended to disappear through MRI examination. In addition, the lesions on the left side were remarkably reduced and the ranges of the frontal and parietal lobes and semi-oval area were reduced. The lesion on the cervical spinal cord also became lighter. B. Patient 2, MRI results revealed that the high signal intensity was reduced at the site next to the left ventricle and basal ganglia. C. Patient in the control group, high signal intensity was visible in the white matter lateral to the bilateral ventricles and semi-oval area. Multiple nodular lesions were observed in the corona radiata and the site next to the lateral ventricles.

Figure 5

Quantitative real time PCR analyses of mRNA of CD86, IL-2, IL-17c, Foxp3, CTLA-4, HLA-DRB1, TGF-β1, and TGF-β2 genes in peripheral blood from patients were shown. Data were normalized to corresponding GAPDH expressions as internal control. The results of mRNA expressions are expressed as fold of control. Evaluation of changes of cytokines with mRNA level in Patient 1 (A-H) and Patient 2 (I-P). The mRNA levels of cytokines were monitored before each transplantation, and compared with their relative levels at the beginning of the trail, which was considered as the control.