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Abstract
Purpose
The aim of study was to compare the differentiation capacity of mesenchymal stem cells (MSCs) obtained from human bone marrow (BM) according to the age of the donors.
Materials and Methods
MSCs were isolated from the BM of young (n=16, 12.5±5.8 years) and elder (n=4, 48.5±7.2 years) patients with the consent of them. We analyzed the cell morphology and the cell surface markers of the MSCs. In addition, we assessed the cell senescence with serial cultures from both age groups. Cell pluripotentiality was analyzed by osteogenic, chondrogenic, and adipogenic induction media. We performed RT-PCR, a measurement of expression of alkaline phosphatase, and staining with von Kossa, safranin O, and oil red O stain.
Results
All of the MSC samples tested, irrespective of the age of the donors, MSCs were all successfully isolated from twenty bone marrows. However, the number of cells of from the young donors was five times greater than that of the elderly donors. Senescence was observed over 10 passages in both age groups. The immunophenotypes of both age groups showed similar patterns. MSCs obtained from young and older donors showed the potential to differentiate into osteogenic, chondrogenic, and adipogenic lineages with no difference for both age groups.
Figures and Tables
Fig. 1
Photomicrographs showing the adherent cells obtained from human bone marrow cells. (A) Adherent cells at day 1. (B) Heterogeneous adherent cells at day 3. (C) Confluent fibroblast-like adherent cells at day 7. An homogeneous population of bipolar spindle-shaped cells was obtained. All representative examples are shown at 100×magnification.
Fig. 2
Osteogenic differentiation of human BM MSCs. (A) Osteogenic induction was detected by alkaline phosphatase expression. (B) Calcium matrix mineralization was detected by the use of von Kossa stain. (C) Expression of mRNA of osteocalcin, alkaline phosphase, osteopontin, Cbfa-1 and collagen type I during osteogenic differentiation. All representative examples are shown at 100×magnification. 0 week: without osteogenic differentiation, 3 weeks: with osteogenic differentiation.
Fig. 3
Chondrogenic differentiation of human BM MSCs. (A) Proteoglycan synthesis was visualized by the use of safranin O. (B) Expression of mRNA of GAPDH, Aggrecan, Collgen type IX, Sox 9, and Collagen type II during chondrogenic differentiation. All representative examples are shown at 400×magnification. 0 week: without chondrogenic differentiation, 3 weeks: with chondrogenic differentiation.
Fig. 4
Adipogenic differentiation of human BM MSCs. (A) Adipogenic induction was apparent by the accumulation of lipid vacuoles within cells, which were visualized by staining with oil red O. (B) Expression of mRNA of PPARγ, Leptin, LPL, C/EBPα, and αP2 during adipogenic differntiaiton. All representative examples are shown at 200×magnification. 0 week: without adipogenic differentiation, 3 weeks: with adipogenic differentiation.
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