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Stem Cells Int. 2019 Apr 4;2019:9271595. doi: 10.1155/2019/9271595. eCollection 2019.

Study on the Dynamic Biological Characteristics of Human Bone Marrow Mesenchymal Stem Cell Senescence.

Chen X1,2, Wang L2,3, Hou J2,3, Li J2,3, Chen L2,3, Xia J2,3, Wang Z2,3, Xiao M2,3, Wang Y2,3.

Author information

1
Department of Anatomy and Histology and Embryology, Basic Medical College, Chengdu University of Traditional Chinese Medicine, 1166 Liutaida Road, Wenjiang District, Chengdu, Sichuan 610075, China.
2
Laboratory of Stem Cells and Tissue Engineering, Chongqing Medical University, 1 Yixueyuan Road, YuZhong District, Chongqing 400016, China.
3
Department of Histology and Embryology, Chongqing Medical University, 1 Yixueyuan Road, Yuzhong District, Chongqing 400016, China.

Abstract

Objective:

To preliminary explore the senescent dynamic changes of the bone marrow mesenchymal stem cells (BMMSCs) by human ageing and its possible mechanism.

Methods:

The bone marrows were harvested from healthy volunteers, and according to volunteers' age, these were divided into group A (≤25 years), group B (26-45 years), group C (46-65 years), and group D (>65 years). Totally, the bone marrows were extracted from the posterior superior iliac spine from volunteers under aseptic conditions. Diluted with isovolumic PBS, followed by centrifugation at 1 × 105/cm2, cells were cultured in a 5% CO2 incubator at 37°C. After three passages, surface marker identification of hBMMSCs was tested by flow cytometry (FCM), oil red O staining was used to observe the ability of osteogenic differentiation, alkaline phosphatase (ALP) staining and the levels of osteocalcin (OST) in the supernatants were used to observe the ability of adipogenic differentiation, senescence-associated β-galactosidase (SA-β-Gal) staining was used to detect the senescent BMSCs, the ability of BMSC proliferation was detected by cell counting kit-8 (CCK-8), the distribution of the cell cycle was analyzed by flow cytometry (FCM), and malondialdehyde (MDA) content, total glutathione peroxidase, total antioxidant capacity, and total superoxide dismutase (SOD) activity was analyzed using enzymatic assay.

Results:

The BMSCs highly expressed CD73 and CD90, but lowly expressed CD34 and CD19/CD14. With age, osteogenic differentiation was markedly increased and audiogenic differentiation was significantly decreased. The number of SA-β-gal-positive cells was significantly increased, the proliferation ability of hBMMSCs declined, the BMSCs were held in the G1 phase, the MDA level of BMSCs was significantly increased, and total glutathione peroxidase, total antioxidant capacity, and SOD activity significantly declined.

Conclusions:

With age, the aging BMSCs were intensified; the mechanism may be related to oxidative damage mediated aging-related pathways.

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