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. 2015;37(2):16.
doi: 10.1007/s11357-014-9743-z. Epub 2015 Feb 19.

Effect of Aged Bone Marrow Microenvironment on Mesenchymal Stem Cell Migration

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Free PMC article

Effect of Aged Bone Marrow Microenvironment on Mesenchymal Stem Cell Migration

Yan-Mei Yang et al. Age (Dordr). .
Free PMC article

Abstract

Mesenchymal stem cells (MSCs) are known to have many notable features, especially their multiple differentiation ability and immunoregulatory capacity. MSCs are important stem cells in the bone marrow (BM), and their characteristics are affected by the BM microenvironment. However, effects of the BM microenvironment on the properties of MSCs are not well understood. In this study, we found that BM from aged mice decreased MSC colony formation. Flow cytometry data showed that the proportion of B220(+) cells in BM from aged mice was significantly lower than that in BM from young mice, while the proportion of CD11b(+), CD3(+), Gr-1(+), or F4/80(+) cells are on the contrary. CD11b(+), B220(+), and Ter119(+) cells from aged mice were not the subsets that decreased MSC colony formation. We further demonstrated that both BM from aged mice and young mice exhibited similar effects on the proliferation of murine MSC cell line C3H10T1/2. However, when cocultured with BM from aged mice, C3H10T1/2 showed slower migration ability. In addition, we found that phosphorylation of JNK (c-Jun N-terminal kinases) in C3H10T1/2 cocultured with BM from aged mice was lower than that in C3H10T1/2 cocultured with BM from young mice. Collectively, our data revealed that BM from aged mice could decrease the migration of MSCs from their niche through regulating the JNK pathway.

Figures

Fig. 1
Fig. 1
Comparison of MSC colony numbers. a Three methods [flushing-out bone marrow (BMA), collagenase-digested bone fragments (BA), bone marrow plus bone fragments (BM A+B A)] were used to isolate and culture the aged mice MSCs. The colony number in the (BM A+B A) group was significantly less than that in the BA group at 6 and 8 days.*P < 0.05. b The method of collagenase-digested bone fragments was used to isolate and culture aged and young mice MSCs. Colonies from aged mice was significantly less than from young mice (BA vs BY), **P < 0.01. c BMA and BMY co-culture with the same bone fragments, *P < 0.05. d Transwell was added to separate bone marrow and bone fragments in culture. BMA decreased the formation of MSCs colonies, *P < 0.05. Data are shown from one of three repeated experiments. (B Y bone fragments from young mice, B A bone fragments from aged mice, BM Y BM from young mice, BM A BM from aged mice, BM+B BM cocultured with B directly, BM/B BM in transwell chamber co-cultured with B)
Fig. 2
Fig. 2
Flow cytometry analysis of BMA and BMY. The proportion of subsets in BMA and BMY: CD3+, CD11b+, Ter119+, Gr1+, and F4/80+ cells in BMA are significantly higher than those in BMY, while B220+ cells in BMA are significantly lower than in BMY. Data are shown from one of three repeated experiments
Fig. 3
Fig. 3
MSC colony formation. Morphology of MSC colonies from different subsets of BMA cocultured with bone fragments at day 3 and day 12. Data are shown from one of three repeated experiments (scale bar = 100 μm)
Fig. 4
Fig. 4
MSC colony number of different subsets of BMA cocultured with bone fragments. a The MSC colony number of B220+, CD11b+, and Ter119+ cells of BMA cocultured with bone fragments are significantly more than the number of BMA co-cultured with bone fragments at day 3.*P < 0.05;**P < 0.01. b The MSC colony number of B220+, CD11b+, and Ter119+ cells of BMA cocultured with bone fragments are more than the number of BMA co-cultured with bone fragments at day 6.*P < 0.05; **P < 0.01. Data are shown from one of three repeated experiments
Fig. 5
Fig. 5
Proliferation and migration of C3H10T1/2 undergoing BMA or BMY stimulation. a Growth curve of C3H10T1/2 cocultured with BMA or BMY. b Wound healing of C3H10T1/2 under BMA or BMY stimulation. (A, B × 40; C, D × 100). Data are shown from one of three repeated experiments (scale bar = 100 μm)
Fig. 6
Fig. 6
Expression of p-p38, p-JNK, p-ERK1/2, and ERK1/2 was examined by Western blot. p38, JNK, and ERK1/2 pathways were all activated, but only p-JNK in the BMY group was higher than in the BMA group at 15, 30, and 60 min. Data are shown from one of three repeated experiments

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