Generation and properties of a new human ventral mesencephalic neural stem cell line

Villa, Ana; Liste, Isabel; Courtois, Elise T.; Seiz, Emma G.; Ramos, Milagros; Meyer, Morten; Juliusson, Bengt; Kusk, Philip; Martínez‐Serrano, Alberto

doi:10.1016/j.yexcr.2009.03.011

Cited by 46 publications

(83 citation statements)

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“…We recently reported on a newly derived human fetal ventral mesencephalic stem cell line, hVMl, that generates high fractions of dopaminergic cells, even after considerable expansion (>10 passages) (Villa et al, 2009). The hVMl cell line was subjected to thorough biochemical, genetic, and immunocytochemical characterization to document its usefulness as a model for developing clinically competent cell lines for PD (Villa et al, 2009).…”

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“…The hVMl cell line was subjected to thorough biochemical, genetic, and immunocytochemical characterization to document its usefulness as a model for developing clinically competent cell lines for PD (Villa et al, 2009). Here we have explored the functional properties of hVMl cells in vitro, based on calcium imaging and electrophysiological investigations.…”

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“…The derivation of the hVMl cell line was previously described in detail (Villa et al, 2009). Briefly, cells were isolated from the ventral mesencephalic region of a dead, aborted human fetus, aged 10 weeks, at Lund University Hospital, according to guidelines approved by the Lund/Malmó Ethical Committee.…”

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“…Briefly, cells were isolated from the ventral mesencephalic region of a dead, aborted human fetus, aged 10 weeks, at Lund University Hospital, according to guidelines approved by the Lund/Malmó Ethical Committee. Cells were immortalized by infection with a retroviral vector coding for v-myc (LTR-vmyc-SV40p-Neo-LTR) (Villa et al, 2000) generating the hVMl polyclonal cell line (Villa et al, 2009). …”

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“…Cells were expanded as attached cultures on poly-L-lysine (PLL, 10 ug/ml)-coated dishes in N2 supplemented DMEM/F12 medium in the presence of 20 ng/ml of each of EGF and bFGF (Villa et al, 2009). To induce differentiation of the cells, bFGF and EGF were withdrawn from the medium and human recombinant glial cell line-derived neurotrophic factor (GDNF, 2 ng/ml, Peprotech) and dibutyryl-cAMP (1 uM, Sigma) were added (Lotharius et al, 2002;Riaz et al, 2002).…”

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Functional properties of the human ventral mesencephalic neural stem cell line hVM1

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Ramos

et al. 2010

Experimental Neurology

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The human fetal ventral mesencephalon-derived stem cell line, hVTVIl, yields high number of tyrosine hydroxylase-expressing presumed dopaminergic neurons upon in vitro differentiation. Here we report that cells generated from this line differentiate into a neuronal phenotype, express electrophysiological properties of functional neurons and respond to neurotransmitters in vitro. However, the electrophysiological properties are immature and the cells require longer maturation time than possible under in vitro

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Functional properties of the human ventral mesencephalic neural stem cell line hVM1

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Seiz

Ramos

et al. 2010

Experimental Neurology

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Pyrolytic Carbon Nanograss Enhances Neurogenesis and Dopaminergic Differentiation of Human Midbrain Neural Stem Cells

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García-López

Heiskanen

et al. 2020

Adv Healthcare Materials

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Advancements in research on the interaction of human neural stem cells (hNSCs) with nanotopographies and biomaterials are enhancing the ability to influence cell migration, proliferation, gene expression, and tailored differentiation toward desired phenotypes. Here, the fabrication of pyrolytic carbon nanograss (CNG) nanotopographies is reported and demonstrated that these can be employed as cell substrates boosting hNSCs differentiation into dopaminergic neurons (DAn), a long-time pursued goal in regenerative medicine based on cell replacement. In the near future, such structures can play a crucial role in the near future for stem-cell based cell replacement therapy (CRT) and bio-implants for Parkinson's disease (PD). The unique combination of randomly distributed nanograss topographies and biocompatible pyrolytic carbon material is optimized to provide suitable mechano-material cues for hNSCs adhesion, division, and DAn differentiation of midbrain hNSCs. The results show that in the presence of the biocoating poly-L-lysine (PLL), the CNG enhances hNSCs neurogenesis up to 2.3-fold and DAn differentiation up to 3.5-fold. Moreover, for the first time, consistent evidence is provided, that CNGs without any PLL coating are not only supporting cell survival but also lead to significantly enhanced neurogenesis and promote hNSCs to acquire dopaminergic phenotype compared to PLL coated topographies.

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Leaky Optoelectrical Fiber for Optogenetic Stimulation and Electrochemical Detection of Dopamine Exocytosis from Human Dopaminergic Neurons

et al. 2019

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In Parkinson's disease, the degeneration of dopaminergic neurons in substantia nigra leads to a decrease in the physiological levels of dopamine in striatum. The existing dopaminergic therapies effectively alleviate the symptoms, albeit they do not revert the disease progression and result in significant adverse effects. Transplanting dopaminergic neurons derived from stem cells could restore dopamine levels without additional motor complications. However, the transplanted cells disperse in vivo and it is not possible to stimulate them on demand to modulate dopamine release to prevent dyskinesia. In order to address these issues, this paper presents a multifunctional leaky optoelectrical fiber for potential neuromodulation and as a cell substrate for application in combined optogenetic stem cell therapy. Pyrolytic carbon coated optical fibers are laser ablated to pattern micro‐optical windows to permit light leakage over a large area. The pyrolytic carbon acts as an excellent electrode for the electrochemical detection of dopamine. Human neural stem cells are genetically modified to express the light sensitive opsin channelrhodopsin‐2 and are differentiated into dopaminergic neurons on the leaky optoelectrical fiber. Finally, light leaking from the micro‐optical windows is used to stimulate the dopaminergic neurons resulting in the release of dopamine that is detected in real‐time using chronoamperometry.

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Generation and properties of a new human ventral mesencephalic neural stem cell line

Cited by 46 publications

References 57 publications

Functional properties of the human ventral mesencephalic neural stem cell line hVM1

Functional properties of the human ventral mesencephalic neural stem cell line hVM1

Pyrolytic Carbon Nanograss Enhances Neurogenesis and Dopaminergic Differentiation of Human Midbrain Neural Stem Cells

Leaky Optoelectrical Fiber for Optogenetic Stimulation and Electrochemical Detection of Dopamine Exocytosis from Human Dopaminergic Neurons

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