Dopaminergic phenotype of hNT cells in vitro

Zigova, Tanja; Barroso, Luis F.; Willing, Alison E.; Saporta, Samuel; McGrogan, Michael; Freeman, Thomas B.; Sanberg, Paul R.

doi:10.1016/s0165-3806(00)00055-9

Cited by 52 publications

(32 citation statements)

References 18 publications

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“…Such non-neuronal expression of ␤-III tubulin is consistent with observations of expression of this tubulin isoform in other human cell types, including astrocytomas (Katsetos et al, 1993(Katsetos et al, , 1998Svendsen et al, 2001). Additional confirmation that ␤-III tubulin was expressed by non-neuronal cells was provided by induction experiments in which A2B5 ϩ cells were grown in the presence of RA and NT-3, a condition that promotes neuronal differentiation and maturation (Henion and Weston, 1994;Ghosh and Greenberg, 1995;Wilkinson et al, 1996;MayerProschel et al, 1997;Zigova et al, 2000). The expression of glia markers under these conditions was retained in ␤-III tubulin-labeled cells, and we did not see any further increase of ␤-III tubulin expression, nor did we detect any other of the mentioned neuronal markers or the appearance of the neuronal morphologies.…”

Section: Discussionsupporting

confidence: 80%

Characterization of A2B5⁺ glial precursor cells from cryopreserved human fetal brain progenitor cells

Dietrich

Noble

Mayer-Pröschel

2002

Glia

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The identification and characterization of human neural precursor cells are critical in extending our understanding of central nervous system development from model animal systems to our own species. Moreover, availability of well-characterized populations of human cells is of potential value in endeavors ranging from cell transplantation to drug screening. We have isolated a population of continuously dividing glial-restricted precursor cells from commercially available cryopreserved 18-20 weeks old fetal brain neural progenitor cells. These human glial-restricted precursor cells are A2B5(+) and do not express polysialylated E-NCAM (PSA-NCAM). They can be grown as purified populations in serum-free medium supplemented with basic fibroblast growth factor (bFGF) and can be induced to generate cells with the antigenic characteristics of oligodendrocytes and distinct astrocytic populations.

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Section: Discussionsupporting

confidence: 80%

Characterization of A2B5⁺ glial precursor cells from cryopreserved human fetal brain progenitor cells

Dietrich

Noble

Mayer-Pröschel

2002

Glia

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“…We have been unable to observe any markers for a dopaminergic phenotype in our cultures, despite our attempts to induce this phenotype via lithium application (Zigova et al 1999(Zigova et al , 2000 or the application of a mix of FGF1 and various co-activators (Iacovitti et al 2001).…”

Section: Neurotransmittersmentioning

confidence: 85%

“…Previous reports showed differentiation into dopaminergic NT2 neurons by exposure either to lithium (Zigova et al 1999(Zigova et al , 2000 or to a mix of growth factors, dopamine, and high intracellular cAMP levels by the application of Fig. 6 Inverted immunofluorescence images of NT2-neurons stained for neurotransmitters (a, c, e, g, i, k) and the corresponding DAPIlabeled nuclei (b, d, f, h, j, l).…”

Section: Neuronal Phenotypementioning

confidence: 99%

Cellular phenotypes of human model neurons (NT2) after differentiation in aggregate culture

Podrygajlo¹,

Tegenge²,

Gierse³

et al. 2009

Cell Tissue Res

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The well-characterized human teratocarcinoma line Ntera2 (NT2) can be differentiated into mature neurons. We have significantly shortened the time-consuming process for generating postmitotic neurons to approximately 4 weeks by introducing a differentiation protocol for free-floating cell aggregates and a subsequent purification step. Here, we characterize the neurochemical phenotypes of the neurons derived from this cell aggregate method. During differentiation, the NT2 cells lose immunoreactivity for vimentin and nestin filaments, which are characteristic for the immature state of neuronal precursors. Instead, they acquire typical neuronal markers such as beta-tubulin type III, microtubule-associated protein 2, and phosphorylated tau, but no astrocyte markers such as glial fibrillary acidic protein. They grow neural processes that express punctate immunoreactivity for synapsin and synaptotagmin suggesting the formation of presynaptic structures. Despite their common clonal origin, neurons cultured for 2-4 weeks in vitro comprise a heterogeneous population expressing several neurotransmitter phenotypes. Approximately 40% of the neurons display glutamatergic markers. A minority of neurons is immunoreactive for serotonin, gamma-amino-butyric acid, and its synthesizing enzyme glutamic acid decarboxylase. We have found no evidence for a dopaminergic phenotype. Subgroups of NT2 neurons respond to the application of nitric oxide donors with the synthesis of cGMP. A major subset shows immunoreactivity to the cholinergic markers choline acetyl-transferase, vesicular acetylcholine transporter, and the non-phosphorylated form of neurofilament H, all indicative of motor neurons. The NT2 system may thus be well suited for research related to motor neuron diseases.

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“…In contrast, hNT-DA neurons are treated for only 4 weeks with retinoic acid after replating and mitotic inhibitor treatment. A shorter treatment time of NT2 cultures with retinoic acid enhances the number of TH-expressing hNT neurons (Zigova et al, 2000a). hNT-DA/Li neurons are hNT-DA neurons that were cotreated with lithium chloride (LiCl) and mitotic inhibitors for 6 days.…”

mentioning

confidence: 99%

Long distance selective fiber outgrowth of transplanted hNT neurons in white matter tracts of the adult rat brain

Baker

Mendez

2005

J of Comparative Neurology

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Terminally differentiated neurons derived from a human teratocarcinoma cell line (NT2N or hNT neurons) are promising as a cell source for transplantation, as they have been shown to be safe for transplantation in humans. We have shown previously that hNT neurons can express a catecholaminergic phenotype in a rat Parkinson model. In this study, we investigated the long-term survival and ability of hNT neurons to express tyrosine hydroxylase and reconstruct the dopamine-denervated nigrostriatal pathway. Hemiparkinsonian rats received grafts of 400,000 viable hNT neurons into each of the denervated striatum and substantia nigra. Robust hNT grafts were detected up to 24 weeks posttransplantation, although few cells expressed tyrosine hydroxylase. Many hNT fibers were often associated with ipsilateral and contralateral white matter tracts--corpus callosum, rostral migratory stream, optic tract, and external capsule. Fewer fibers were associated with the superior cerebellar peduncle, medial lemniscus, and nigrostriatal pathway. Axons also projected into the frontal cortex and extended parallel to the surface of the brain in the superficial cortical layers. These pathways were seen in all grafted animals, suggesting that specific guidance cues exist in the adult brain governing hNT fiber outgrowth. Injured adult axons and transplanted embryonic neuronal axons rarely extend for such distances in the adult nervous system. We propose that elucidating the factors promoting and guiding hNT axonal outgrowth could provide important clues to enhancing regeneration and target reinnervation in the adult brain, two factors of critical importance for cell restoration strategies aimed at brain repair.

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Dopaminergic phenotype of hNT cells in vitro

Cited by 52 publications

References 18 publications

Characterization of A2B5⁺ glial precursor cells from cryopreserved human fetal brain progenitor cells

Characterization of A2B5⁺ glial precursor cells from cryopreserved human fetal brain progenitor cells

Cellular phenotypes of human model neurons (NT2) after differentiation in aggregate culture

Long distance selective fiber outgrowth of transplanted hNT neurons in white matter tracts of the adult rat brain

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Dopaminergic phenotype of hNT cells in vitro

Cited by 52 publications

References 18 publications

Characterization of A2B5+ glial precursor cells from cryopreserved human fetal brain progenitor cells

Characterization of A2B5+ glial precursor cells from cryopreserved human fetal brain progenitor cells

Cellular phenotypes of human model neurons (NT2) after differentiation in aggregate culture

Long distance selective fiber outgrowth of transplanted hNT neurons in white matter tracts of the adult rat brain

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Characterization of A2B5⁺ glial precursor cells from cryopreserved human fetal brain progenitor cells

Characterization of A2B5⁺ glial precursor cells from cryopreserved human fetal brain progenitor cells