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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

Indirect co-culture with BM-MSCs results in enhanced differentiation of Gba−/− NSCs into specific cell types. (A) Differentiation of NSCs was induced for 7 days, with or without BM-MSC co-culture, using an indirect three-dimensional system. Cells were stained with specific cell type markers. Representative fluorescence images and quantitative analysis of βIII-tubulin/DAPI (B, C), MBP/DAPI (D), and GFAP/DAPI (E) are shown (scale bar, 50 μm, n = 5 per group). Data are presented as the mean percentage of immunopositive cells to DAPI-stained cells for each group. (C) Total outgrowth of differentiated neuron dendrites and the number of dendritic processes are shown. For each measurement, at least 60 cells per disc were counted from randomly selected fields of view (n = 5 discs from different mouse origins). All data are presented as the mean ± SEM. *p < 0.05, ***p < 0.005.
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f3-molce-38-9-806: Indirect co-culture with BM-MSCs results in enhanced differentiation of Gba−/− NSCs into specific cell types. (A) Differentiation of NSCs was induced for 7 days, with or without BM-MSC co-culture, using an indirect three-dimensional system. Cells were stained with specific cell type markers. Representative fluorescence images and quantitative analysis of βIII-tubulin/DAPI (B, C), MBP/DAPI (D), and GFAP/DAPI (E) are shown (scale bar, 50 μm, n = 5 per group). Data are presented as the mean percentage of immunopositive cells to DAPI-stained cells for each group. (C) Total outgrowth of differentiated neuron dendrites and the number of dendritic processes are shown. For each measurement, at least 60 cells per disc were counted from randomly selected fields of view (n = 5 discs from different mouse origins). All data are presented as the mean ± SEM. *p < 0.05, ***p < 0.005.

Mentions: In addition to their ability for self-renewal, the second hallmark of stemness in NSCs is multilineage differentiation (Reynolds and Weiss, 1996). We dissociated NSs and co-cultured cells with BM-MSCs in differentiation media. Immunocytochemistry was performed to investigate the effects of BM-MSCs on Gba−/− NSC cultures and their capacity for differentiation into the three main neural lineages (Fig. 3A). Expression of three neural lineage markers (βIII-tubulin, MBP and GFAP) were detected in NSC-derived cells regardless of whether they were co-cultured with BM-MSCs (Figs. 3B–3E). In comparison with Gba+/− cells, the efficiency of neuronal differentiation was not significantly different. We observed significantly reduced neurite outgrowth and number of processes in differentiated neurons from Gba−/− NSC cultures (Figs. 3B and 3C). Indirect co-culture of Gba−/− NSCs with BM-MSCs resulted in significantly enhanced efficiency of neuronal differentiation, increased neurite outgrowth, and an increased number of processes (Fig. 3C). Compared with Gba+/− NSCs, we observed a reduced rate of differentiation for MBP-expressing cells in Gba−/− NSC cultures (Fig. 3D). Co-culture with BM-MSCs resulted in a significant increase in the number of differentiating oligodendro- cytes in Gba−/− NSC cultures (Fig. 3D). Compared with Gba+/− cells however, there were no significant changes in the numbers of GFAP-positive cells in differentiated Gba−/− NSC cultures (Fig. 3E). Overall, our results indicate that BM-MSCs selectively promote neurogenesis, neuronal morphogenesis, and oligo- genesis of 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)

Indirect co-culture with BM-MSCs results in enhanced differentiation of Gba−/− NSCs into specific cell types. (A) Differentiation of NSCs was induced for 7 days, with or without BM-MSC co-culture, using an indirect three-dimensional system. Cells were stained with specific cell type markers. Representative fluorescence images and quantitative analysis of βIII-tubulin/DAPI (B, C), MBP/DAPI (D), and GFAP/DAPI (E) are shown (scale bar, 50 μm, n = 5 per group). Data are presented as the mean percentage of immunopositive cells to DAPI-stained cells for each group. (C) Total outgrowth of differentiated neuron dendrites and the number of dendritic processes are shown. For each measurement, at least 60 cells per disc were counted from randomly selected fields of view (n = 5 discs from different mouse origins). All data are presented as the mean ± SEM. *p < 0.05, ***p < 0.005.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4588724&req=5

f3-molce-38-9-806: Indirect co-culture with BM-MSCs results in enhanced differentiation of Gba−/− NSCs into specific cell types. (A) Differentiation of NSCs was induced for 7 days, with or without BM-MSC co-culture, using an indirect three-dimensional system. Cells were stained with specific cell type markers. Representative fluorescence images and quantitative analysis of βIII-tubulin/DAPI (B, C), MBP/DAPI (D), and GFAP/DAPI (E) are shown (scale bar, 50 μm, n = 5 per group). Data are presented as the mean percentage of immunopositive cells to DAPI-stained cells for each group. (C) Total outgrowth of differentiated neuron dendrites and the number of dendritic processes are shown. For each measurement, at least 60 cells per disc were counted from randomly selected fields of view (n = 5 discs from different mouse origins). All data are presented as the mean ± SEM. *p < 0.05, ***p < 0.005.
Mentions: In addition to their ability for self-renewal, the second hallmark of stemness in NSCs is multilineage differentiation (Reynolds and Weiss, 1996). We dissociated NSs and co-cultured cells with BM-MSCs in differentiation media. Immunocytochemistry was performed to investigate the effects of BM-MSCs on Gba−/− NSC cultures and their capacity for differentiation into the three main neural lineages (Fig. 3A). Expression of three neural lineage markers (βIII-tubulin, MBP and GFAP) were detected in NSC-derived cells regardless of whether they were co-cultured with BM-MSCs (Figs. 3B–3E). In comparison with Gba+/− cells, the efficiency of neuronal differentiation was not significantly different. We observed significantly reduced neurite outgrowth and number of processes in differentiated neurons from Gba−/− NSC cultures (Figs. 3B and 3C). Indirect co-culture of Gba−/− NSCs with BM-MSCs resulted in significantly enhanced efficiency of neuronal differentiation, increased neurite outgrowth, and an increased number of processes (Fig. 3C). Compared with Gba+/− NSCs, we observed a reduced rate of differentiation for MBP-expressing cells in Gba−/− NSC cultures (Fig. 3D). Co-culture with BM-MSCs resulted in a significant increase in the number of differentiating oligodendro- cytes in Gba−/− NSC cultures (Fig. 3D). Compared with Gba+/− cells however, there were no significant changes in the numbers of GFAP-positive cells in differentiated Gba−/− NSC cultures (Fig. 3E). Overall, our results indicate that BM-MSCs selectively promote neurogenesis, neuronal morphogenesis, and oligo- genesis of 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