In vitro growth and differentiation of canine olfactory bulb-derived neural progenitor cells under variable culture conditions

Walton, Raquel M.; Wolfe, John H.

doi:10.1016/j.jneumeth.2007.12.011

Cited by 16 publications

(21 citation statements)

References 71 publications

(89 reference statements)

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“…F 0479) secondary antibody and remained for one hour in a dark container, at room temperature. All slides were washed three times with DPBS before mounting in Prolong containing Hoechst 33342.This protocol with adaptations was described by Walton and Wolfe 16 .…”

Section: Neuronal Differentiation and Immunofluorescence Protocolmentioning

confidence: 99%

Rabbit olfactory stem cells. Isolation protocol and characterization

Ercolin

Roballo²,

Casals³

et al. 2016

Acta Cir. Bras.

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PURPOSE:To describe a new technique for isolation of a mesenchymal stem cells (MSCs) population from the olfactory mucosa in rabbits. METHODS:Olfactory stem cells (OSCs) were retrieved from under the cribriform plate of the Ethmoid bone. Several assays were accomplished to characterize the cell population and attest its viability in vitro. The cells were submitted to flow cytometry with the antibodies CD34, CD45, CD73, CD79, CD90 and CD105 and also they were induced to differentiate in three lineages. Functional evaluation involved analysis of in vitro growth behavior, colony forming unit like fibroblasts (CFU-f) and cryopreservation response.Further transduction with Green Fluorescent Protein (GFP) was also performed. RESULTS:The OSCs showed mesenchymal features, as positive response to CD34, CD73 and CD90 antibodies and plasticity.Additionally, these cells have high proliferated rate, and they could be cultured through many passages and kept the ability to proliferate and differentiate after cryopreservation. The positive response to the transduction signalizes the possibility of cellular tracking in vivo. This is a desirable feature in case those cells are used for pre-clinical trials. CONCLUSION:The cells harvested were mesenchymal stem cells and the technique described is therefore efficient for rabbit olfactory stem cells isolation.

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Section: Neuronal Differentiation and Immunofluorescence Protocolmentioning

confidence: 99%

Rabbit olfactory stem cells. Isolation protocol and characterization

Ercolin

Roballo²,

Casals³

et al. 2016

Acta Cir. Bras.

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“…Neural progenitor cells (NPCs) have now been derived from the brain or spinal cord of mammalian species, including mouse (Vukicevic et al 2010;Lindstrom et al 2009;Hernandez-Benitez et al 2010), dog (Walton and Wolfe 2008;Milward et al 1997), rat (Go et al 2009), human (Luo et al 2010;Liu et al 2010) and pig (Schwartz et al 2005). PNPCs seems to be less immunogenic and thus survive better than porcine neural xenografts (Armstrong et al 2001;Barker et al 2000), up to 5 months in rats (Harrower et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Spontaneous differentiation of porcine neural progenitors in vitro

Yin

Guo

et al. 2011

Cytotechnology

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The pig is the non-primate species that is immunologically closest to humans, and has been considered as an alternative source to human allografts for transplantation. In fact, there has been recent interest in identifying and culturing porcine neural progenitor cells (PNPCs) in vitro, but the long-term culturing has not yet been characterized. Here, we reported the spontaneous differentiation of PNPCs into neuronal and glial cells. For in vitro cultures, the primary cells of the subventricular zone of the forebrain striatum were cultured in the presence of epidermal growth factor and basic fibroblast growth factor to allow the growth of spherical masses that exhibit sustained growth and self-renewal capacity. After growth factor removal, the neurospheres with 10 and 130 days of culture spontaneously differentiated into Tuj1-positive neurons and GFAP-positive astrocytes as seen by double immunocytofluorescence. Molecular characterization using reverse transcription-polymerase chain reaction showed that neurospheres expressed nestin, neuron-specific enolase, and glial fibrillary acidic protein (GFAP). In addition, after cultured in the differentiation medium for 3 months, the growth of neurosphere became slow and displayed cystic structures with the same morphology as that of embryonic bodies derived from embryonic stem cells. It is concluded that PNPCs have the ability to provide an expandable source of neural cells that can develop into neuronal and glial subtypes.

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“…They can be expanded ex vivo manyfold and are able to differentiate in vitro into neurons and glia (38, 46). Moreover, the use of cNPCs derived from the postnatal OB affords the opportunity for autologous transplantation.…”

Section: Discussionmentioning

confidence: 99%

“…Newly isolated canine OB-NPCs (OB-cNPCs) were plated into 25-cm 2 tissue culture flasks (Corning, Acton, MA) coated with 10 μg/mL poly-D-lysine (Sigma) and expanded, as reported previously (38). Briefly, cells were plated at a concentration of 4 × 10 4 /cm 2 in 10% serum-containing medium consisting of Dulbecco modified Eagle medium: F12 (1:1 ratio; GibcoBRL Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum, 1% N2 supplement (GibcoBRL), 1% penicillin/streptomycin/fungizone (100 U/mL penicillin, 100 μg/mL streptomycin, 0.25 μg/mL amphotericin B; GibcoBRL), and 1% L-glutamine (2 mM; GibcoBRL) with 20 ng/mL epidermal growth factor (recombinant murine; Roche, Nutley, NJ), 20 ng/mL basic fibroblast growth factor (recombinant human; Promega, Madison, WI), and heparin (5 μg/mL; Sigma).…”

Section: Methodsmentioning

confidence: 99%

Transplantation and Magnetic Resonance Imaging of Canine Neural Progenitor Cell Grafts in the Postnatal Dog Brain

Walton

Magnitsky

Seiler

et al. 2008

J Neuropathol Exp Neurol

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Cellular transplantation in the form of bone marrow has been one of the primary treatments of many lysosomal storage diseases (LSDs). Although bone marrow transplantation can help central nervous system manifestations in some cases, it has little impact in many LSD patients. Canine models of neurogenetic LSDs provide the opportunity for modeling central nervous system transplantation strategies in brains that more closely approximate the size and architectural complexity of the brains of children. Canine olfactory bulb-derived neural progenitor cells (NPCs) isolated from dog brains were expanded ex vivo and implanted into the caudate nucleus/thalamus or cortex of allogeneic dogs. Canine olfactory bulb-derived NPCs labeled with micron-sized superparamagnetic iron oxide particles were detected by magnetic resonance imaging both in vivo and postmortem. Grafts expressed markers of NPCs (i.e. nestin and glial fibrillary acidic protein), but not the neuronal markers Map2ab or β-tubulin III. The NPCs were from dogs with the LSD mucopolysaccharidosis VII, which is caused by a deficiency of β-glucuronidase. When mucopolysaccharidosis VII canine olfactory bulb-NPCs that were genetically corrected with a lentivirus vector ex vivo were transplanted into mucopolysaccharidosis VII recipient brains, they were detected histologically by β-glucuronidase expression in areas identified by antemortem magnetic resonance imaging tracking. These results demonstrate the potential for ex vivo stem cell-based gene therapy and noninvasive tracking of therapeutic grafts in vivo.

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In vitro growth and differentiation of canine olfactory bulb-derived neural progenitor cells under variable culture conditions

Cited by 16 publications

References 71 publications

Rabbit olfactory stem cells. Isolation protocol and characterization

Rabbit olfactory stem cells. Isolation protocol and characterization

Spontaneous differentiation of porcine neural progenitors in vitro

Transplantation and Magnetic Resonance Imaging of Canine Neural Progenitor Cell Grafts in the Postnatal Dog Brain

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