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The isolation, differentiation, and survival in vivo of multipotent cells from the postnatal rat filum terminale.

Jha RM, Chrenek R, Magnotti LM, Cardozo DL - PLoS ONE (2013)

Bottom Line: Neurospheres derived from the rat FT are amenable to in vitro expansion in the presence of a combination of growth factors.Through directed differentiation using sonic hedgehog and retinoic acid in combination with various neurotrophic factors, FT-derived neurospheres generated motor neurons that were capable of forming neuromuscular junctions in vitro.In addition, FT-derived progenitors that were injected into chick embryos survived and could differentiate into both neurons and glia in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT
Neural stem cells (NSCs) are undifferentiated cells in the central nervous system (CNS) that are capable of self-renewal and can be induced to differentiate into neurons and glia. Current sources of mammalian NSCs are confined to regions of the CNS that are critical to normal function and surgically difficult to access, which limits their therapeutic potential in human disease. We have found that the filum terminale (FT), a previously unexplored, expendable, and easily accessible tissue at the caudal end of the spinal cord, is a source of multipotent cells in postnatal rats and humans. In this study, we used a rat model to isolate and characterize the potential of these cells. Neurospheres derived from the rat FT are amenable to in vitro expansion in the presence of a combination of growth factors. These proliferating, FT-derived cells formed neurospheres that could be induced to differentiate into neural progenitor cells, neurons, astrocytes, and oligodendrocytes by exposure to serum and/or adhesive substrates. Through directed differentiation using sonic hedgehog and retinoic acid in combination with various neurotrophic factors, FT-derived neurospheres generated motor neurons that were capable of forming neuromuscular junctions in vitro. In addition, FT-derived progenitors that were injected into chick embryos survived and could differentiate into both neurons and glia in vivo.

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Related in: MedlinePlus

Variable expression of MNR2 in FT-derived neurospheres undergoing MN differentiation.Scatter graph illustrating the variability in MNR2 expression in differentiated neurospheres under four sets of differentiation conditions. The X axis is the experiment number, and the Y axis is the average proportion of cells derived from each neurosphere expressing MNR2, Tuj-1 and/or GFAP. MNR2, which was used to identify MN generation, was expressed by cells in roughly the same proportion among conditions (1), (2), and (3) with more inconsistent results under condition (4). In 3 of the 28 experiments where the small molecule agonist, Hh-Ag1.3, was used in place of Shh-N, 100% expression of MNR2 was observed. See Results for more detail.
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pone-0065974-g008: Variable expression of MNR2 in FT-derived neurospheres undergoing MN differentiation.Scatter graph illustrating the variability in MNR2 expression in differentiated neurospheres under four sets of differentiation conditions. The X axis is the experiment number, and the Y axis is the average proportion of cells derived from each neurosphere expressing MNR2, Tuj-1 and/or GFAP. MNR2, which was used to identify MN generation, was expressed by cells in roughly the same proportion among conditions (1), (2), and (3) with more inconsistent results under condition (4). In 3 of the 28 experiments where the small molecule agonist, Hh-Ag1.3, was used in place of Shh-N, 100% expression of MNR2 was observed. See Results for more detail.

Mentions: While neurospheres treated with Shh-N gave rise to differentiated neurons, only 20–40% of these neurons expressed MN markers such as MNR2, Isl1, Lim3 and ChAT (n = 14). Increasing the Shh-N concentration from 400 to 1000 nM did not appear to alter the outcome. When we instead used the hedgehog agonist Hh-Ag1.3 (n = 9, 1.5 µM), 95–100% of the differentiated neurons expressed MN markers (Figure 8). This suggests that at these concentrations, the agonist may be more effective than the actual peptide in generating MNs from FT-derived neurospheres.


The isolation, differentiation, and survival in vivo of multipotent cells from the postnatal rat filum terminale.

Jha RM, Chrenek R, Magnotti LM, Cardozo DL - PLoS ONE (2013)

Variable expression of MNR2 in FT-derived neurospheres undergoing MN differentiation.Scatter graph illustrating the variability in MNR2 expression in differentiated neurospheres under four sets of differentiation conditions. The X axis is the experiment number, and the Y axis is the average proportion of cells derived from each neurosphere expressing MNR2, Tuj-1 and/or GFAP. MNR2, which was used to identify MN generation, was expressed by cells in roughly the same proportion among conditions (1), (2), and (3) with more inconsistent results under condition (4). In 3 of the 28 experiments where the small molecule agonist, Hh-Ag1.3, was used in place of Shh-N, 100% expression of MNR2 was observed. See Results for more detail.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3675200&req=5

pone-0065974-g008: Variable expression of MNR2 in FT-derived neurospheres undergoing MN differentiation.Scatter graph illustrating the variability in MNR2 expression in differentiated neurospheres under four sets of differentiation conditions. The X axis is the experiment number, and the Y axis is the average proportion of cells derived from each neurosphere expressing MNR2, Tuj-1 and/or GFAP. MNR2, which was used to identify MN generation, was expressed by cells in roughly the same proportion among conditions (1), (2), and (3) with more inconsistent results under condition (4). In 3 of the 28 experiments where the small molecule agonist, Hh-Ag1.3, was used in place of Shh-N, 100% expression of MNR2 was observed. See Results for more detail.
Mentions: While neurospheres treated with Shh-N gave rise to differentiated neurons, only 20–40% of these neurons expressed MN markers such as MNR2, Isl1, Lim3 and ChAT (n = 14). Increasing the Shh-N concentration from 400 to 1000 nM did not appear to alter the outcome. When we instead used the hedgehog agonist Hh-Ag1.3 (n = 9, 1.5 µM), 95–100% of the differentiated neurons expressed MN markers (Figure 8). This suggests that at these concentrations, the agonist may be more effective than the actual peptide in generating MNs from FT-derived neurospheres.

Bottom Line: Neurospheres derived from the rat FT are amenable to in vitro expansion in the presence of a combination of growth factors.Through directed differentiation using sonic hedgehog and retinoic acid in combination with various neurotrophic factors, FT-derived neurospheres generated motor neurons that were capable of forming neuromuscular junctions in vitro.In addition, FT-derived progenitors that were injected into chick embryos survived and could differentiate into both neurons and glia in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT
Neural stem cells (NSCs) are undifferentiated cells in the central nervous system (CNS) that are capable of self-renewal and can be induced to differentiate into neurons and glia. Current sources of mammalian NSCs are confined to regions of the CNS that are critical to normal function and surgically difficult to access, which limits their therapeutic potential in human disease. We have found that the filum terminale (FT), a previously unexplored, expendable, and easily accessible tissue at the caudal end of the spinal cord, is a source of multipotent cells in postnatal rats and humans. In this study, we used a rat model to isolate and characterize the potential of these cells. Neurospheres derived from the rat FT are amenable to in vitro expansion in the presence of a combination of growth factors. These proliferating, FT-derived cells formed neurospheres that could be induced to differentiate into neural progenitor cells, neurons, astrocytes, and oligodendrocytes by exposure to serum and/or adhesive substrates. Through directed differentiation using sonic hedgehog and retinoic acid in combination with various neurotrophic factors, FT-derived neurospheres generated motor neurons that were capable of forming neuromuscular junctions in vitro. In addition, FT-derived progenitors that were injected into chick embryos survived and could differentiate into both neurons and glia in vivo.

Show MeSH
Related in: MedlinePlus