Introduction
Materials and methods
Isolation and expansion of mesenchymal stem cells
Flow cytometric characterization and differentiation of mesenchymal stem cells
Anti-CD3-induced cell proliferation
Measurement of in vivoT-cell proliferation
Quantitative polymerase chain reaction
Bio-Plex protein array system
Collagen-induced arthritis induction and treatment protocols
Measurement of anti-CII antibodies and delayed-type hypersensitivity to CII
Statistical analysis
Results
Generation of mesenchymal stem cells and phenotypical analysis
The isolated mesenchymal stem cells differentiate into osteogenic and adipogenic lineages
Mesenchymal stem cells suppress anti-CD3-induced T-cell proliferation in vitroby a mechanism involving interferon-gamma, inducible nitric oxide, and cyclo-oxigenase-2
Mesenchymal stem cell treatment has no effect on the development of collagen-induced arthritis
Experiment | Routeb | Administration timec | Treatmentd | Cumulative incidencee, fraction (percentage) | Score of arthritis, mean ± SEMe, f |
---|---|---|---|---|---|
1 | i.v. | Day -1 | Control | 4/8 (50%) | 2.0 ± 0.9 |
Wild-type MSCs | 1/2 (50%) | 1.5 ± 1.5 | |||
IFN-γR KO MSCs | 3/7 (43%) | 1.6 ± 1.0 | |||
Treg cells | 2/8 (25%) | 0.4 ± 0.3 | |||
2 | i.v. | Day 16 | Control | 4/10 (40%) | 1.3 ± 0.7 |
Wild-type MSCs | 6/8 (75%) | 2.1 ± 0.6 | |||
IFN-γR KO MSCs | 4/10 (40%) | 0.8 ± 0.4 | |||
3 | i.v. | Days 16 and 23 | Control | 5/10 (50%) | 3.0 ± 1.4 |
Wild-type MSCs | 5/9 (56%) | 2.5 ± 0.9 | |||
IFN-γR KO MSCs | 5/10 (50%) | 2.0 ± 0.8 | |||
4 | i.p. | Days 16, 23, and 30 | Control | 7/8 (88%) | 5.1 ± 1.5 |
Wild-type MSCs | 6/7 (86%) | 5.3 ± 1.2 | |||
IFN-γR KO MSCs | 8/9 (89%) | 4.6 ± 1.6 | |||
5 | i.p. | Days 16, 23, and 30 | Control | 7/11 (64%) | 1.3 ± 0.5 |
C57BL/6 MSCs | 8/11 (73%) | 2.7 ± 1.2 |
Discussion
MSC source | MSC administration | |||||||
---|---|---|---|---|---|---|---|---|
Reference | Organisma | Strainb | Organc | Transfectiond | Routee | Dosef | Timeg | Resulth |
[26] | Mouse | C3 | Cell line | n.a. | i.v. | 1 × 106 | 0 | 0 |
21 | 0 | |||||||
4 × 106 | 0 | 0 | ||||||
21 | - | |||||||
IL-10 | i.v. | 1 × 106 | 0 | 0 | ||||
[23] | Mouse | C57Bl/10 | BM | n.a. | i.p. | 5 × 106 | 0 | + |
21 | + | |||||||
[27] | Mouse | DBA/1 | BM | n.a. | i.v. | 1 × 106 | 21+28+35 | 0 |
IL-10 | i.v. | 1 × 106 | 21+28+35 | + | ||||
[24] | Human | n.a. | Adipose | n.a. | i.p. | 1 × 106 | 5 a.d.o. | + |
Mouse | C57Bl/6 | Adipose | n.a. | i.p. | 1 × 106 | 5 a.d.o. | + | |
Mouse | DBA/1 | Adipose | n.a. | i.p. | 1 × 106 | 5 a.d.o. | + | |
[25] | Rat | n.s. | BM | n.a. | i.v. | 2 × 106 | 1 a.d.o. | + |