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.
Highlights
Bone marrow is the most important hematopoietic organ and is composed of hematopoietic cells and bone marrow mesenchymal stem cells (BMMSCs) at different stages of development [1]
The results demonstrated that the cultured cells were typical Human bone marrow mesenchymal stem cells (hBMMSCs) (Figure 1)
Bone marrow mesenchymal cells have been considered as a good support function for sustaining hematopoietic stem cells (HSCs), and they have been regarded as a promising seed due to their capacity to differentiate into osteoblasts and so forth [4]
Summary
Bone marrow is the most important hematopoietic organ and is composed of hematopoietic cells and bone marrow mesenchymal stem cells (BMMSCs) at different stages of development [1]. BMMSCs are the main component of the hematopoietic inductive microenvironment (HIM), where hematopoietic stem cells (HSCs) lodge, proliferate, differentiate, and are regulated, and they can reflect the functional status of a hematopoietic inductive microenvironment and maintain the steady state [2]. In the hematopoietic inductive microenvironment, hematopoietic stem cells generate specific progenitor cells and the self-renewal divisions necessary to sustain themselves; those functions all associate with HIM. It has been observed for a long time that with advancing age, the number of HSCs decreases, which leads to deficiency. As the core component of HIM, as to whether BMMSCs can manifest age-related senescence like HSCs can, there have been few reports concerning this. To better elucidate the underlying mechanism(s) in age-associated BMMSCs, here we investigated the effects of age in BMMSCs by humans
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