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A method to generate human mesenchymal stem cell-derived neurons which express and are excited by multiple neurotransmitters.

Greco SJ, Zhou C, Ye JH, Rameshwar P - Biol Proced Online (2008)

Bottom Line: MSCs are bone marrow (BM)-derived cells which undergo lineage- specific differentiation to generate bone, fat, cartilage and muscle, but are also capable of transdifferentiating into defined ectodermal and endodermal tissues.Our neuronal protocol utilizes freshly harvested human MSCs cultured on specific surfaces and exposed to an induction cocktail consisting of low serum concentration, retinoic acid (RA), growth factors and supplements.Here we report on the types of neurotransmitters produced by the neurons, and demonstrate that the cells are electrically responsive to exogenous neurotransmitter administration.

View Article: PubMed Central - HTML - PubMed

Affiliation: Graduate School of Biomedical Sciences, UMDNJ-New Jersey Medical School, MSB, Rm. E-579 185 South Orange Ave, Newark, NJ 07103, USA.

ABSTRACT
The present study describes a protocol to generate heterogenous populations of neurotransmitter-producing neurons from human mesenchymal stem cells (MSCs). MSCs are bone marrow (BM)-derived cells which undergo lineage- specific differentiation to generate bone, fat, cartilage and muscle, but are also capable of transdifferentiating into defined ectodermal and endodermal tissues. The purpose of this study is to evaluate the potential of MSCs as an alternative source of customized neurons for experimental neurobiology or other regenerative approaches. Our neuronal protocol utilizes freshly harvested human MSCs cultured on specific surfaces and exposed to an induction cocktail consisting of low serum concentration, retinoic acid (RA), growth factors and supplements. Here we report on the types of neurotransmitters produced by the neurons, and demonstrate that the cells are electrically responsive to exogenous neurotransmitter administration.

No MeSH data available.


Colocalization of SP and GAD in induced MSCs.D12 cells were co-labeled with FITC-anti-GAD and PE-anti-SP, and then counter-labeled with DAPI. Figure shows representative labelings of five different experiments. Images are shown at 40X magnification
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Figure 03: Colocalization of SP and GAD in induced MSCs.D12 cells were co-labeled with FITC-anti-GAD and PE-anti-SP, and then counter-labeled with DAPI. Figure shows representative labelings of five different experiments. Images are shown at 40X magnification

Mentions: The next set of studies asked whether the peptidergic (SP+) and putative GABAergic (GAD+) neurons existed as distinct sub-populations, or whether some cells were capable of producing both neurotransmitters. To address this question, we co-labeled D12 induced MSCs for GAD (green) and SP (red) expression and observed the cells by immunofluorescence (Fig. 3). Very few cells (<1%) were found to co-express SP and GAD (Merge –far right panel). The results suggest that the peptidergic and putative GABAergic cells primarily exist as two distinct sub-populations.


A method to generate human mesenchymal stem cell-derived neurons which express and are excited by multiple neurotransmitters.

Greco SJ, Zhou C, Ye JH, Rameshwar P - Biol Proced Online (2008)

Colocalization of SP and GAD in induced MSCs.D12 cells were co-labeled with FITC-anti-GAD and PE-anti-SP, and then counter-labeled with DAPI. Figure shows representative labelings of five different experiments. Images are shown at 40X magnification
© Copyright Policy - open acces
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2683550&req=5

Figure 03: Colocalization of SP and GAD in induced MSCs.D12 cells were co-labeled with FITC-anti-GAD and PE-anti-SP, and then counter-labeled with DAPI. Figure shows representative labelings of five different experiments. Images are shown at 40X magnification
Mentions: The next set of studies asked whether the peptidergic (SP+) and putative GABAergic (GAD+) neurons existed as distinct sub-populations, or whether some cells were capable of producing both neurotransmitters. To address this question, we co-labeled D12 induced MSCs for GAD (green) and SP (red) expression and observed the cells by immunofluorescence (Fig. 3). Very few cells (<1%) were found to co-express SP and GAD (Merge –far right panel). The results suggest that the peptidergic and putative GABAergic cells primarily exist as two distinct sub-populations.

Bottom Line: MSCs are bone marrow (BM)-derived cells which undergo lineage- specific differentiation to generate bone, fat, cartilage and muscle, but are also capable of transdifferentiating into defined ectodermal and endodermal tissues.Our neuronal protocol utilizes freshly harvested human MSCs cultured on specific surfaces and exposed to an induction cocktail consisting of low serum concentration, retinoic acid (RA), growth factors and supplements.Here we report on the types of neurotransmitters produced by the neurons, and demonstrate that the cells are electrically responsive to exogenous neurotransmitter administration.

View Article: PubMed Central - HTML - PubMed

Affiliation: Graduate School of Biomedical Sciences, UMDNJ-New Jersey Medical School, MSB, Rm. E-579 185 South Orange Ave, Newark, NJ 07103, USA.

ABSTRACT
The present study describes a protocol to generate heterogenous populations of neurotransmitter-producing neurons from human mesenchymal stem cells (MSCs). MSCs are bone marrow (BM)-derived cells which undergo lineage- specific differentiation to generate bone, fat, cartilage and muscle, but are also capable of transdifferentiating into defined ectodermal and endodermal tissues. The purpose of this study is to evaluate the potential of MSCs as an alternative source of customized neurons for experimental neurobiology or other regenerative approaches. Our neuronal protocol utilizes freshly harvested human MSCs cultured on specific surfaces and exposed to an induction cocktail consisting of low serum concentration, retinoic acid (RA), growth factors and supplements. Here we report on the types of neurotransmitters produced by the neurons, and demonstrate that the cells are electrically responsive to exogenous neurotransmitter administration.

No MeSH data available.