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Defective Self-Renewal and Differentiation of GBA-Deficient Neural Stem Cells Can Be Restored By Macrophage Colony-Stimulating Factor.

Lee H, Bae JS, Jin HK - Mol. Cells (2015)

Bottom Line: We found that neural stem cells (NSCs) derived from a neuronopathic GD model exhibited decreased ability for self-renewal and neuronal differentiation.Enhanced proliferation and neuronal differentiation of GBA-deficient NSCs was associated with elevated release of macrophage colony-stimulating factor (M-CSF) from BM-MSCs.Our findings suggest that soluble M-CSF derived from BM-MSCs can modulate GBA-deficient NSCs, resulting in their improved proliferation and neuronal differentiation.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell Neuroplasticity Research Group, Cell and Matrix Research Institute, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Korea.

ABSTRACT
Gaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the glucocerebrosidase gene (GBA), which encodes the lysosomal enzyme glucosylceramidase (GCase). Deficiency in GCase leads to characteristic visceral pathology and lethal neurological manifestations in some patients. Investigations into neurogenesis have suggested that neurodegenerative disorders, such as GD, could be overcome or at least ameliorated by the generation of new neurons. Bone marrow-derived mesenchymal stem cells (BM-MSCs) are potential candidates for use in the treatment of neurodegenerative disorders because of their ability to promote neurogenesis. Our objective was to examine the mechanism of neurogenesis by BM-MSCs in GD. We found that neural stem cells (NSCs) derived from a neuronopathic GD model exhibited decreased ability for self-renewal and neuronal differentiation. Co-culture of GBA-deficient NSCs with BM-MSCs resulted in an enhanced capacity for self-renewal, and an increased ability for differentiation into neurons or oligodendrocytes. Enhanced proliferation and neuronal differentiation of GBA-deficient NSCs was associated with elevated release of macrophage colony-stimulating factor (M-CSF) from BM-MSCs. Our findings suggest that soluble M-CSF derived from BM-MSCs can modulate GBA-deficient NSCs, resulting in their improved proliferation and neuronal differentiation.

No MeSH data available.


Related in: MedlinePlus

BM-MSCs enhance the self-renewal and proliferation of Gba−/− NSCs. (A) Experimental design to determine the effects of BM-MSCs on Gba−/− NSCs. (B) Light micro-graphs of NSs following BM-MSC co-culture with Gba−/− NSCs (scale bar, 1 mm). Self-renewal capacity is expressed as the number of NSs (n = 6 per group). (C) Effect of BM-MSCs on the proliferation of Gba−/− NSCs (scale bar, 20 μm). Proliferation ability was assessed by the percentage of BrdU-positive cells (n = 7 per group). (D) Effect of BM-MSCs on the GCase activity of Gba−/− NSCs (n = 3 per group). All data are presented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.005.
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f2-molce-38-9-806: BM-MSCs enhance the self-renewal and proliferation of Gba−/− NSCs. (A) Experimental design to determine the effects of BM-MSCs on Gba−/− NSCs. (B) Light micro-graphs of NSs following BM-MSC co-culture with Gba−/− NSCs (scale bar, 1 mm). Self-renewal capacity is expressed as the number of NSs (n = 6 per group). (C) Effect of BM-MSCs on the proliferation of Gba−/− NSCs (scale bar, 20 μm). Proliferation ability was assessed by the percentage of BrdU-positive cells (n = 7 per group). (D) Effect of BM-MSCs on the GCase activity of Gba−/− NSCs (n = 3 per group). All data are presented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.005.

Mentions: To examine the neurogenic potential of BM-MSCs, we used self-renewal and proliferation assays (Fig. 2A). The number of NSs in Gba−/− cultures was significantly lower compared with that in Gba+/− cultures (Fig. 2B). In addition, indirect co-culture of Gba−/− NSCs with BM-MSCs resulted in the increased formation of NSs (Fig. 2B). BM-MSCs also induced an increase in NS formation in Gba+/− cultures. To assess proliferation, adherent NSCs were pulsed with BrdU and the proportion of labeled cells analyzed (Fig. 2C). BM-MSCs exhibited increased BrdU incorporation in Gba+/− and Gba−/− cultures in comparison with untreated control cultures (Fig. 2C). These results indicate that BM-MSCs promote NSC proliferation. To validate whether the effect of BM-MSCs was mediated by GCase activity, we analyzed its activity in NSCs co-cultured with BM-MSCs. Activities of GCase in Gba−/− and Gba+/− cells co-cultured with NSCs were similar to those in cells that were not co-cultured with BM-MSCs (Fig. 2D). Our results suggest that BM-MSCs increase self-renewal and proliferation of NSCs. Although the effects of BM-MSCs were not limited to conditions where GCase activity was knocked down, our findings indicate that BM-MSCs promote self-renewal and proliferation of Gba−/− NSCs. In addition, these results provide evidence that the effects of BM-MSCs can be attributed to the indirect effect of neurogenic soluble factors secreted from BM-MSCs.


Defective Self-Renewal and Differentiation of GBA-Deficient Neural Stem Cells Can Be Restored By Macrophage Colony-Stimulating Factor.

Lee H, Bae JS, Jin HK - Mol. Cells (2015)

BM-MSCs enhance the self-renewal and proliferation of Gba−/− NSCs. (A) Experimental design to determine the effects of BM-MSCs on Gba−/− NSCs. (B) Light micro-graphs of NSs following BM-MSC co-culture with Gba−/− NSCs (scale bar, 1 mm). Self-renewal capacity is expressed as the number of NSs (n = 6 per group). (C) Effect of BM-MSCs on the proliferation of Gba−/− NSCs (scale bar, 20 μm). Proliferation ability was assessed by the percentage of BrdU-positive cells (n = 7 per group). (D) Effect of BM-MSCs on the GCase activity of Gba−/− NSCs (n = 3 per group). All data are presented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.005.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4588724&req=5

f2-molce-38-9-806: BM-MSCs enhance the self-renewal and proliferation of Gba−/− NSCs. (A) Experimental design to determine the effects of BM-MSCs on Gba−/− NSCs. (B) Light micro-graphs of NSs following BM-MSC co-culture with Gba−/− NSCs (scale bar, 1 mm). Self-renewal capacity is expressed as the number of NSs (n = 6 per group). (C) Effect of BM-MSCs on the proliferation of Gba−/− NSCs (scale bar, 20 μm). Proliferation ability was assessed by the percentage of BrdU-positive cells (n = 7 per group). (D) Effect of BM-MSCs on the GCase activity of Gba−/− NSCs (n = 3 per group). All data are presented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.005.
Mentions: To examine the neurogenic potential of BM-MSCs, we used self-renewal and proliferation assays (Fig. 2A). The number of NSs in Gba−/− cultures was significantly lower compared with that in Gba+/− cultures (Fig. 2B). In addition, indirect co-culture of Gba−/− NSCs with BM-MSCs resulted in the increased formation of NSs (Fig. 2B). BM-MSCs also induced an increase in NS formation in Gba+/− cultures. To assess proliferation, adherent NSCs were pulsed with BrdU and the proportion of labeled cells analyzed (Fig. 2C). BM-MSCs exhibited increased BrdU incorporation in Gba+/− and Gba−/− cultures in comparison with untreated control cultures (Fig. 2C). These results indicate that BM-MSCs promote NSC proliferation. To validate whether the effect of BM-MSCs was mediated by GCase activity, we analyzed its activity in NSCs co-cultured with BM-MSCs. Activities of GCase in Gba−/− and Gba+/− cells co-cultured with NSCs were similar to those in cells that were not co-cultured with BM-MSCs (Fig. 2D). Our results suggest that BM-MSCs increase self-renewal and proliferation of NSCs. Although the effects of BM-MSCs were not limited to conditions where GCase activity was knocked down, our findings indicate that BM-MSCs promote self-renewal and proliferation of Gba−/− NSCs. In addition, these results provide evidence that the effects of BM-MSCs can be attributed to the indirect effect of neurogenic soluble factors secreted from BM-MSCs.

Bottom Line: We found that neural stem cells (NSCs) derived from a neuronopathic GD model exhibited decreased ability for self-renewal and neuronal differentiation.Enhanced proliferation and neuronal differentiation of GBA-deficient NSCs was associated with elevated release of macrophage colony-stimulating factor (M-CSF) from BM-MSCs.Our findings suggest that soluble M-CSF derived from BM-MSCs can modulate GBA-deficient NSCs, resulting in their improved proliferation and neuronal differentiation.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell Neuroplasticity Research Group, Cell and Matrix Research Institute, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Korea.

ABSTRACT
Gaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the glucocerebrosidase gene (GBA), which encodes the lysosomal enzyme glucosylceramidase (GCase). Deficiency in GCase leads to characteristic visceral pathology and lethal neurological manifestations in some patients. Investigations into neurogenesis have suggested that neurodegenerative disorders, such as GD, could be overcome or at least ameliorated by the generation of new neurons. Bone marrow-derived mesenchymal stem cells (BM-MSCs) are potential candidates for use in the treatment of neurodegenerative disorders because of their ability to promote neurogenesis. Our objective was to examine the mechanism of neurogenesis by BM-MSCs in GD. We found that neural stem cells (NSCs) derived from a neuronopathic GD model exhibited decreased ability for self-renewal and neuronal differentiation. Co-culture of GBA-deficient NSCs with BM-MSCs resulted in an enhanced capacity for self-renewal, and an increased ability for differentiation into neurons or oligodendrocytes. Enhanced proliferation and neuronal differentiation of GBA-deficient NSCs was associated with elevated release of macrophage colony-stimulating factor (M-CSF) from BM-MSCs. Our findings suggest that soluble M-CSF derived from BM-MSCs can modulate GBA-deficient NSCs, resulting in their improved proliferation and neuronal differentiation.

No MeSH data available.


Related in: MedlinePlus