J Korean Soc Endocrinol.  2003 Jun;18(3):296-305.

The Effects of Aging on the Proliferation and Differentiation of Osteoblasts from Human Mesenchymal Stem Cells

Affiliations
  • 1Department of Internal Medicine, The Catholic University of Korea, College of Medicine.
  • 2Department of Neurosurgery, The Catholic University of Korea, College of Medicine.
  • 3Department of Orthopedic Surgery, The Catholic University of Korea, College of Medicine.
  • 4Hemopoietic Stem Cell Transplantation Center, The Catholic University of Korea, College of Medicine.
  • 5Mizmedi hospital.
  • 6Department of Internal Medicine, Sungkyunkwan University School of Medicine, Seoul, Korea.

Abstract

BACKGROUND: Osteoblasts originate from osteoprogenitor cells in bone marrow stroma, termed mesenchymal stem cells (MSCs) or bone marrow stromal cells. Each MSC forms colonies (colony forming units-fibroblasts [CFU-Fs]) when cultured ex vivo. There are some reports about the age-related changes of the number and osteogenic potential of osteoprogenitor cells, but any relationship has not been clearly established in humans. In this study, we counted MSCs using CFU-Fs count and examined the proliferative capacity and differentiation potential of osteoprogenitor cells. Finally, we analyzed how these parameters varied with donor age.
METHODS
Bone marrow was obtained from the iliac crest of young (n=6, 27.2+/-8.6 years old) and old (n=10, 57.4+/-6.7 years old) healthy donors. Mononuclear cells, including MSCs, were isolated and cultured in osteogenic medium. In primary culture, we compared the colony-forming efficiency of MSCs between the two groups and determined the matrix calcification. When primary culture showed near confluence, the cells were subcultured. Alkaline phosphatase activity, osteocalcinexpression by RT-PCR and proliferative potential by MTT assay were examined by the time course of secondary culture.
RESULTS
At the 15th day of primary culture, the mean number of CFU-Fs was significantly higher in the younger donors (young: 148.3+/-28.9, old: 54.3+/-9.1, p=0.02) and the mean size of CFU-Fs was also larger in the younger donors than the older donors. However, matrix calcification was not different between the two groups (young: 103.6+/-50.6, old: 114.0+/-56.5, p=NS). In secondary culture, alkaline phosphatase activities were significantly lower in the older donors. The younger donors showed peak alkaline phosphatase activity at day 10, while the older donors didn't showed a remarkable peak (young: 935.5+/-115.0U/mg, old: 578.4+/-115.7U/mg, p<0.05). Total cell number as a proliferative index increased progressively during the secondary culture and a significantly greater cell number was noted in the younger donors. Osteocalcin expression was generally upregulated in the younger donors, but this was not statistically significant.
CONCLUSION
Our study shows that the number of osteoprogenitor cells is decreased during aging and that the proliferative capacity and differentiation potential of osteoprogenitor cells seem to be reduced during aging.


MeSH Terms

Aging*
Alkaline Phosphatase
Bone Marrow
Cell Count
Humans*
Insulin Resistance
Mesenchymal Stromal Cells*
Osteoblasts*
Osteocalcin
Tissue Donors
Alkaline Phosphatase
Osteocalcin
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