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Novel stem/progenitor cells with neuronal differentiation potential reside in the leptomeningeal niche.

Bifari F, Decimo I, Chiamulera C, Bersan E, Malpeli G, Johansson J, Lisi V, Bonetti B, Fumagalli G, Pizzolo G, Krampera M - J. Cell. Mol. Med. (2009)

Bottom Line: Once injected into the adult brain, these cells survive and differentiate into neurons, thus showing that their neuronal differentiation potential is operational also in vivo.In conclusion, our data provide evidence that a specific population of immature cells endowed of neuronal differentiation potential is resident in the leptomeninges throughout the life.As leptomeninges cover the entire central nervous system, these findings could have relevant implications for studies on cortical development and for regenerative medicine applied to neurological disorders.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Experimental Medicine, Stem Cell Research Laboratory, Section of Hematology, University of Verona, Verona, Italy.

ABSTRACT
Stem cells capable of generating neural differentiated cells are recognized by the expression of nestin and reside in specific regions of the brain, namely, hippocampus, subventricular zone and olfactory bulb. For other brain structures, such as leptomeninges, which contribute to the correct cortex development and functions, there is no evidence so far that they may contain stem/precursor cells. In this work, we show for the first time that nestin-positive cells are present in rat leptomeninges during development up to adulthood. The newly identified nestin-positive cells can be extracted and expanded in vitro both as neurospheres, displaying high similarity with subventricular zone-derived neural stem cells, and as homogeneous cell population with stem cell features. In vitro expanded stem cell population can differentiate with high efficiency into excitable cells with neuronal phenotype and morphology. Once injected into the adult brain, these cells survive and differentiate into neurons, thus showing that their neuronal differentiation potential is operational also in vivo. In conclusion, our data provide evidence that a specific population of immature cells endowed of neuronal differentiation potential is resident in the leptomeninges throughout the life. As leptomeninges cover the entire central nervous system, these findings could have relevant implications for studies on cortical development and for regenerative medicine applied to neurological disorders.

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In vitro neural differentiation of leptomeningeal cells after adherent culture expansion (A) Real-time gene expression analysis of Nestin and Mtap2 (MAP2) in leptomeninges-derived cells in basal condition and after differentiation. The bars show folds change ± S.D. of normalized mRNA expression levels measured before and after differentiation (P < 0.01). (B, C, D) Differentiated cells, stained with antibodies against MAP2 (red). Arrows in (C) indicate dendritic spines. (D) BrdU staining (green) indicates that the MAP2-positive cells (red) derived from replicating cells. MAP2-positive cells also expressed components of the synaptic apparatus, including the presynaptic marker synaptophysin (E), the glutamate ionotropic receptor sub-unit GluR2 (G) and glutamate decarboxylase (GAD67) (F). Scale bars 50 μm.
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fig06: In vitro neural differentiation of leptomeningeal cells after adherent culture expansion (A) Real-time gene expression analysis of Nestin and Mtap2 (MAP2) in leptomeninges-derived cells in basal condition and after differentiation. The bars show folds change ± S.D. of normalized mRNA expression levels measured before and after differentiation (P < 0.01). (B, C, D) Differentiated cells, stained with antibodies against MAP2 (red). Arrows in (C) indicate dendritic spines. (D) BrdU staining (green) indicates that the MAP2-positive cells (red) derived from replicating cells. MAP2-positive cells also expressed components of the synaptic apparatus, including the presynaptic marker synaptophysin (E), the glutamate ionotropic receptor sub-unit GluR2 (G) and glutamate decarboxylase (GAD67) (F). Scale bars 50 μm.

Mentions: To determine whether the leptomeningeal adherent/expanded cells had neural differentiation potential, they were cultured with differentiating medium for at least 1 month and then analyzed for the presence of the neural marker MAP2. Real-time gene expression analysis of leptomeninges-derived cells in basal condition and after differentiation showed a statistically significant increase in Mtap2 (MAP2) and decrease in nestin expression levels (P < 0.01). Immunofluorescence analysis showed that cells differentiated in cultures into MAP2-positive neurons with high efficiency (30–50% of cells, n= 6 experiments; Fig. 6B–D). Most of MAP2-negative cells were nestin-positive. Some GFAP-positive astrocytes and rare NG2- or O4-positive oligodendrocytic precursors were also found in the differentiated cultures (data not shown). Moreover, MAP2-positive cells showed other features of neuronal phenotype, including distinct neuritic arborization, dendritic spines (arrows in Fig. 6C) and the expression of presynaptic protein synaptophysin (Fig. 6E). A fraction of the differentiated MAP2-positive cells also expressed the GluR2 sub-unit of the ionotropic AMPA-glutamate receptor (Fig. 6G) and the glutamate decarboxylase (GAD67), marker of GABAergic neurons (Fig. 6F).


Novel stem/progenitor cells with neuronal differentiation potential reside in the leptomeningeal niche.

Bifari F, Decimo I, Chiamulera C, Bersan E, Malpeli G, Johansson J, Lisi V, Bonetti B, Fumagalli G, Pizzolo G, Krampera M - J. Cell. Mol. Med. (2009)

In vitro neural differentiation of leptomeningeal cells after adherent culture expansion (A) Real-time gene expression analysis of Nestin and Mtap2 (MAP2) in leptomeninges-derived cells in basal condition and after differentiation. The bars show folds change ± S.D. of normalized mRNA expression levels measured before and after differentiation (P < 0.01). (B, C, D) Differentiated cells, stained with antibodies against MAP2 (red). Arrows in (C) indicate dendritic spines. (D) BrdU staining (green) indicates that the MAP2-positive cells (red) derived from replicating cells. MAP2-positive cells also expressed components of the synaptic apparatus, including the presynaptic marker synaptophysin (E), the glutamate ionotropic receptor sub-unit GluR2 (G) and glutamate decarboxylase (GAD67) (F). Scale bars 50 μm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4516477&req=5

fig06: In vitro neural differentiation of leptomeningeal cells after adherent culture expansion (A) Real-time gene expression analysis of Nestin and Mtap2 (MAP2) in leptomeninges-derived cells in basal condition and after differentiation. The bars show folds change ± S.D. of normalized mRNA expression levels measured before and after differentiation (P < 0.01). (B, C, D) Differentiated cells, stained with antibodies against MAP2 (red). Arrows in (C) indicate dendritic spines. (D) BrdU staining (green) indicates that the MAP2-positive cells (red) derived from replicating cells. MAP2-positive cells also expressed components of the synaptic apparatus, including the presynaptic marker synaptophysin (E), the glutamate ionotropic receptor sub-unit GluR2 (G) and glutamate decarboxylase (GAD67) (F). Scale bars 50 μm.
Mentions: To determine whether the leptomeningeal adherent/expanded cells had neural differentiation potential, they were cultured with differentiating medium for at least 1 month and then analyzed for the presence of the neural marker MAP2. Real-time gene expression analysis of leptomeninges-derived cells in basal condition and after differentiation showed a statistically significant increase in Mtap2 (MAP2) and decrease in nestin expression levels (P < 0.01). Immunofluorescence analysis showed that cells differentiated in cultures into MAP2-positive neurons with high efficiency (30–50% of cells, n= 6 experiments; Fig. 6B–D). Most of MAP2-negative cells were nestin-positive. Some GFAP-positive astrocytes and rare NG2- or O4-positive oligodendrocytic precursors were also found in the differentiated cultures (data not shown). Moreover, MAP2-positive cells showed other features of neuronal phenotype, including distinct neuritic arborization, dendritic spines (arrows in Fig. 6C) and the expression of presynaptic protein synaptophysin (Fig. 6E). A fraction of the differentiated MAP2-positive cells also expressed the GluR2 sub-unit of the ionotropic AMPA-glutamate receptor (Fig. 6G) and the glutamate decarboxylase (GAD67), marker of GABAergic neurons (Fig. 6F).

Bottom Line: Once injected into the adult brain, these cells survive and differentiate into neurons, thus showing that their neuronal differentiation potential is operational also in vivo.In conclusion, our data provide evidence that a specific population of immature cells endowed of neuronal differentiation potential is resident in the leptomeninges throughout the life.As leptomeninges cover the entire central nervous system, these findings could have relevant implications for studies on cortical development and for regenerative medicine applied to neurological disorders.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Experimental Medicine, Stem Cell Research Laboratory, Section of Hematology, University of Verona, Verona, Italy.

ABSTRACT
Stem cells capable of generating neural differentiated cells are recognized by the expression of nestin and reside in specific regions of the brain, namely, hippocampus, subventricular zone and olfactory bulb. For other brain structures, such as leptomeninges, which contribute to the correct cortex development and functions, there is no evidence so far that they may contain stem/precursor cells. In this work, we show for the first time that nestin-positive cells are present in rat leptomeninges during development up to adulthood. The newly identified nestin-positive cells can be extracted and expanded in vitro both as neurospheres, displaying high similarity with subventricular zone-derived neural stem cells, and as homogeneous cell population with stem cell features. In vitro expanded stem cell population can differentiate with high efficiency into excitable cells with neuronal phenotype and morphology. Once injected into the adult brain, these cells survive and differentiate into neurons, thus showing that their neuronal differentiation potential is operational also in vivo. In conclusion, our data provide evidence that a specific population of immature cells endowed of neuronal differentiation potential is resident in the leptomeninges throughout the life. As leptomeninges cover the entire central nervous system, these findings could have relevant implications for studies on cortical development and for regenerative medicine applied to neurological disorders.

Show MeSH
Related in: MedlinePlus