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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

M-CSF enhances the self-renewal, proliferation, and neuronal differentiation of Gba−/− NSCs. (A) Gba−/− NSCs were treated with different concentrations of recombinant murine M-CSF (2-50 ng/ml), and self-renewal was assessed. M-CSF at concentrations of 10 and 50 ng/ml increased the number of Gba−/− NSs (n = 3 per group). (B) M-CSF affects the proliferation of Gba−/− NSCs (scale bar, 20 μm). Proliferation ability was assessed by the percentage of BrdU-positive cells (n = 3 per group). (C) Representative fluorescence images and quantitation of βIII-tubulin (scale bar, 50 μm; n = 3 per group). All data are presented as the mean ± SEM. §p < 0.05, §§§p < 0.005 compared with Gba+/− controls. #p < 0.05 compared with non-treated controls. *p < 0.05, ***p < 0.005.
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f5-molce-38-9-806: M-CSF enhances the self-renewal, proliferation, and neuronal differentiation of Gba−/− NSCs. (A) Gba−/− NSCs were treated with different concentrations of recombinant murine M-CSF (2-50 ng/ml), and self-renewal was assessed. M-CSF at concentrations of 10 and 50 ng/ml increased the number of Gba−/− NSs (n = 3 per group). (B) M-CSF affects the proliferation of Gba−/− NSCs (scale bar, 20 μm). Proliferation ability was assessed by the percentage of BrdU-positive cells (n = 3 per group). (C) Representative fluorescence images and quantitation of βIII-tubulin (scale bar, 50 μm; n = 3 per group). All data are presented as the mean ± SEM. §p < 0.05, §§§p < 0.005 compared with Gba+/− controls. #p < 0.05 compared with non-treated controls. *p < 0.05, ***p < 0.005.

Mentions: To examine the neurogenic potential of M-CSF, we evaluated the effects of recombinant murine M-CSF on the self-renewal of NSs at different concentrations. M-CSF increased the number of NSs in Gba−/− cultures at 10 and 50 ng/ml (Fig. 5A). Therefore, we used 10 ng/ml M-CSF for subsequent experiments. Compared with Gba+/− NSs, the self-renewal ability of Gba−/− NSs was significantly decreased. The Gba−/− NSs treated with M-CSF exhibited increased formation of NSs (Fig. 5A). To assess proliferation, the percentage of BrdU-labeled cells was determined. M-CSF increased BrdU incorporation in Gba−/− cultures relative to that in untreated Gba−/− cultures (Fig. 5B). In addition to examining the effects of M-CSF on neuronal differentiation, NSs were dissociated and treated with M-CSF in differentiation media. After 7 days, the expression of neuronal markers was evaluated in NSC-derived cells. Compared with Gba+/− cells, Gba−/− neurons showed significantly decreased neuronal outgrowth and a reduced number of processes (Fig. 5C). M-CSF treatment of Gba−/− NSCs resulted in increased efficiency of neuronal differentiation, along with an increase in the number of processes and enhanced neuronal outgrowth (Fig. 5C). Taken together, our findings suggest that M-CSF increases self-renewal, proliferation, neuronal differentiation and neuronal morphogenesis of Gba−/− NSCs.


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)

M-CSF enhances the self-renewal, proliferation, and neuronal differentiation of Gba−/− NSCs. (A) Gba−/− NSCs were treated with different concentrations of recombinant murine M-CSF (2-50 ng/ml), and self-renewal was assessed. M-CSF at concentrations of 10 and 50 ng/ml increased the number of Gba−/− NSs (n = 3 per group). (B) M-CSF affects the proliferation of Gba−/− NSCs (scale bar, 20 μm). Proliferation ability was assessed by the percentage of BrdU-positive cells (n = 3 per group). (C) Representative fluorescence images and quantitation of βIII-tubulin (scale bar, 50 μm; n = 3 per group). All data are presented as the mean ± SEM. §p < 0.05, §§§p < 0.005 compared with Gba+/− controls. #p < 0.05 compared with non-treated controls. *p < 0.05, ***p < 0.005.
© Copyright Policy
Related In: Results  -  Collection

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

f5-molce-38-9-806: M-CSF enhances the self-renewal, proliferation, and neuronal differentiation of Gba−/− NSCs. (A) Gba−/− NSCs were treated with different concentrations of recombinant murine M-CSF (2-50 ng/ml), and self-renewal was assessed. M-CSF at concentrations of 10 and 50 ng/ml increased the number of Gba−/− NSs (n = 3 per group). (B) M-CSF affects the proliferation of Gba−/− NSCs (scale bar, 20 μm). Proliferation ability was assessed by the percentage of BrdU-positive cells (n = 3 per group). (C) Representative fluorescence images and quantitation of βIII-tubulin (scale bar, 50 μm; n = 3 per group). All data are presented as the mean ± SEM. §p < 0.05, §§§p < 0.005 compared with Gba+/− controls. #p < 0.05 compared with non-treated controls. *p < 0.05, ***p < 0.005.
Mentions: To examine the neurogenic potential of M-CSF, we evaluated the effects of recombinant murine M-CSF on the self-renewal of NSs at different concentrations. M-CSF increased the number of NSs in Gba−/− cultures at 10 and 50 ng/ml (Fig. 5A). Therefore, we used 10 ng/ml M-CSF for subsequent experiments. Compared with Gba+/− NSs, the self-renewal ability of Gba−/− NSs was significantly decreased. The Gba−/− NSs treated with M-CSF exhibited increased formation of NSs (Fig. 5A). To assess proliferation, the percentage of BrdU-labeled cells was determined. M-CSF increased BrdU incorporation in Gba−/− cultures relative to that in untreated Gba−/− cultures (Fig. 5B). In addition to examining the effects of M-CSF on neuronal differentiation, NSs were dissociated and treated with M-CSF in differentiation media. After 7 days, the expression of neuronal markers was evaluated in NSC-derived cells. Compared with Gba+/− cells, Gba−/− neurons showed significantly decreased neuronal outgrowth and a reduced number of processes (Fig. 5C). M-CSF treatment of Gba−/− NSCs resulted in increased efficiency of neuronal differentiation, along with an increase in the number of processes and enhanced neuronal outgrowth (Fig. 5C). Taken together, our findings suggest that M-CSF increases self-renewal, proliferation, neuronal differentiation and neuronal morphogenesis of Gba−/− NSCs.

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