Equivalent osteoblastic differentiation function of human mesenchymal stem cells from rheumatoid arthritis in comparison with osteoarthritis.
OBJECTIVE
To evaluate the osteoblastic differentiation of human mesenchymal stem cells (hMSCs) in patients with RA.


METHODS
Heparinized bone marrow aspirate was obtained from patients with OA and RA. Mononuclear cells were cultured for 2 weeks and a colony-forming assay was performed. The phenotype of cells was analysed by flow cytometry. Passage 2 cells were cultured with beta-glycerophosphate (bGP) in the control group and bGP, ascorbic acid and dexamethasone in the differentiation group. After 2 weeks, ALP staining and activity were performed. After 3 weeks, Alizarin Red S assay was performed. Total RNA was extracted from cells cultured for 2 and 3 weeks. Gene expression of bone formation factor was examined by real-time PCR.


RESULTS
The phenotype of cells was identical in both OA and RA and the content was thought to be hMSCs. The results of ALP activity and Alizarin Red S assay showed higher levels in the differentiation group for both OA and RA samples compared with the control group. The results of a colony-forming assay were identical in both OA and RA samples. Gene expression in the differentiation group was higher than in the control group in both OA and RA samples. There was no significant difference between OA and RA samples in all experiments.


CONCLUSIONS
The function of osteoblastic differentiation of hMSCs is similar between OA and RA.
Morimoto
Daiki
D
Department of Orthopedics, Osaka University Graduate School of Medicine, Osaka, Japan.
Kuroda
Shoko
S
Kizawa
Takuji
T
Nomura
Koji
K
Higuchi
Chikahisa
C
Yoshikawa
Hideki
H
Tomita
Tetsuya
T
D000368:Aged; D000369:Aged, 80 and over; D000469:Alkaline Phosphatase / Q000032:analysis; D001172:Arthritis, Rheumatoid / Q000473:pathology*; D015415:Biomarkers / Q000032:analysis; D016022:Case-Control Studies; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000473:pathology*; D008875:Middle Aged; D010003:Osteoarthritis / Q000473:pathology*; D010006:Osteoblasts / Q000473:pathology*; D015675:Osteocalcin / Q000032:analysis; D053495:Osteopontin / Q000032:analysis; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013194:Staining and Labeling; D018709:Statistics, Nonparametric
D015415:Biomarkers; D015675:Osteocalcin; D053495:Osteopontin; D000469:Alkaline Phosphatase