<i>In Vitro</i>Generation of Adult Rat Olfactory Sensory Neurons and Regulation of Maturation by Coculture with CNS Tissues

Grill, Raymond J.; Pixley, Sarah K.

doi:10.1523/jneurosci.17-09-03120.1997

Cited by 19 publications

(18 citation statements)

References 44 publications

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“…The majority of previous reports utilize embryonic or neonatal cells (Calof and Chikaraishi, 1989; Chuah et al , 1991; Ronnett et al , 1991; Pixley, 1992; Mumm et al , 1996; Cunningham et al , 1999; McEntire and Pixley, 2000; Carter et al , 2004). Exceptions are found in a few reports describing a culture system of the cells harvested from adult OE (Grill and Pixley, 1997; Liu et al , 1998; Feron et al , 1999; Newman, 2000). …”

Section: Discussionmentioning

confidence: 97%

Maintaining epitheliopoietic potency when culturing olfactory progenitors

Jang

Lambropoulos

Woo

et al. 2008

Experimental Neurology

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The olfactory epithelium is remarkable for the persistence of multipotent, neurocompetent progenitor and stem cells throughout life that can replace all of the various cell types of the epithelium following injury. The therapeutic exploitation of the neurocompetent stem cells of the adult olfactory epithelium would be facilitated by the development of a culture system that maintains the in vivo potency of the progenitors while they are expanded and/or manipulated. We have used an air-liquid interface culture protocol, in which a feeder cell layer of 3T3 cells is established on the underside of a culture insert and FACS-isolated or unsorted progenitor cells from the methyl bromide-lesioned adult rodent epithelium are seeded on upper side. Under these conditions, epithelial cells other than HBCs are capable of organizing themselves into complex 3-dimensional, epithelium-lined spheres, which can be passaged. The spheres contain cells with the molecular phenotype of globose basal cells, horizontal basal cells, sustentacular cells and neurons. Spheres derived from mice that express the green fluorescent protein constitutively can be dissociated after 6 days in vitro and directly transplanted into the epithelium of wild type, methyl bromide-lesioned mice via nasal infusion. The resulting clones contain the various cell types observed in aggregate when globose basal cells are transplanted acutely. In contrast, the same cells cultured as two dimensional, submerged cultures undergo fibroblastic transition after transplantation and do not integrate into the epithelium. In conclusion, the culture system described here maintains the potency of progenitors, which can then participate in epitheliopoiesis in vivo.

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Section: Discussionmentioning

confidence: 97%

Maintaining epitheliopoietic potency when culturing olfactory progenitors

Jang

Lambropoulos

Woo

et al. 2008

Experimental Neurology

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“…Olfactory receptor precursors cultured on cortical astrocytes proliferate extensively and differentiate into mature olfactory receptor neurons (19,20). Cortical neuron precursors from the embryo are maintained in division by contact with astrocytes (14).…”

Section: Discussionmentioning

confidence: 99%

Interaction between astrocytes and adult subventricular zone precursors stimulates neurogenesis

Lim

Alvarez-Buylla

1999

Proc. Natl. Acad. Sci. U.S.A.

328

225

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Neurogenesis continues in the mammalian subventricular zone (SVZ) throughout life. However, the signaling and cell-cell interactions required for adult SVZ neurogenesis are not known. In vivo, migratory neuroblasts (type A cells) and putative precursors (type C cells) are in intimate contact with astrocytes (type B cells). Type B cells also contact each other. We reconstituted SVZ cell-cell interactions in a culture system free of serum or exogenous growth factors. Culturing dissociated postnatal or adult SVZ cells on astrocyte monolayers-but not other substratessupported extensive neurogenesis similar to that observed in vivo. SVZ precursors proliferated rapidly on astrocytes to form colonies containing up to 100 type A neuroblasts. By fractionating the SVZ cell dissociates with differential adhesion to immobilized polylysine, we show that neuronal colonyforming precursors were concentrated in a fraction enriched for type B and C cells. Pure type A cells could migrate in chains but did not give rise to neuronal colonies. Because astrocyteconditioned medium alone was not sufficient to support SVZ neurogenesis, direct cell-cell contact between astrocytes and SVZ neuronal precursors may be necessary for the production of type A cells.Throughout life, the subventricular zone (SVZ) retains a neurogenic population of cells (reviewed in refs. 1-5). Cells born in the neonatal (6) and adult rodent (7) SVZ migrate along a restricted pathway called the rostral migratory stream to the olfactory bulb where they differentiate into interneurons. The SVZ of adult mice lies directly under the ependyma of the lateral wall of the lateral ventricle (8).Three SVZ cell types have been identified based on their ultrastructural and immunocytochemical characteristics (2, 7, 9): (i) type A cells are migratory neuroblasts; (ii) type B cells are astrocytes; and (iii) type C cells are putative precursors. The spatial relationship between these cells is depicted in Fig. 1 Inset. Both type A and type C cells are apposed to the type B astrocytes. Type B cells also have intercellular contacts with each other. Type A cells migrate in chains through glial tubes composed of type B cells, and type C cells are found as small clusters along these chains of migrating neuroblasts. Type A cells are immunopositive for neuronal markers including neuron-specific tubulin (Tuj1) and the polysialylated form of the neural cell-adhesion molecule (PSA-NCAM). Type B cells contain glial-fibrillary acidic protein (GFAP), a marker of astrocytes. Type C cells are ultrastructurally immature and do not stain for either neuronal or glial markers.The type B cells can be classified further: B1 cells contact the ependyma and B2 cells do not. Type A, B2, and C cells are mitotic. Based on their immature characteristics, mitotic activity, and juxtaposition to the chains of type A cells, the type C cells have been proposed to be the immediate precursors of the migratory type A neuroblasts (9).How SVZ neurogenesis is maintained in the adult brain is not well understoo...

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“…Olfactory ensheathing glia (OEG) within the olfactory nerve and bulb nerve layer are also a source of growth factors, including ciliary neurotrophic factor (CNTF), insulin-like growth factor-1 (IGF-1) and fibroblast growth factor-2 (FGF2) (MacKay-Sim and Chuah, 2000). Survival-promoting effects on cultured OSNs have been demonstrated for individual substances within several of these classes of factors, including PDGFs (Newman et al, 2000) and neurotrophins (Holcomb et al, 1995) and co-culture of OSNs with bulb cells similarly promotes their maturation and survival (Grill and Pixley, 1997). …”

Section: Introductionmentioning

confidence: 99%

Long‐term survival of olfactory sensory neurons after target depletion

Sultan-Styne

Toledo

Walker

et al. 2009

J of Comparative Neurology

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Life-long addition and elimination of neurons within the adult olfactory epithelium and olfactory bulb allows for adaptive structural responses to sensory experience, learning, and recovery after injury. The interdependence of the two structures is highlighted by the shortened life span of sensory neurons deprived of bulb contact, and has prompted the hypothesis that trophic cues from the bulb contribute to their survival. The specific identity and source of these signals remain unknown. To investigate the potential role of target neurons in this support, we employed a neurotoxic lesion to selectively remove them while preserving the remaining nerve projection pathway, and examined the dynamics of sensory neuron proliferation and survival. Pulse-labeling of progenitors with bromodeoxyuridine showed that, as with surgical bulb removal, increased apoptosis in the epithelium triggered accelerated production of new neurons after chemical depletion of target cells. Rather than undergoing premature death, a large subpopulation of these neurons survived long term. The combination of increased proliferation and extended survival resulted in essentially normal numbers of new sensory neurons surviving for as long as 5 weeks, with an accompanying restoration of olfactory marker protein expression. Changes in neurotrophic factor expression levels as measured by quantitative polymerase chain reaction (Q-PCR), and in bulb cell populations, including the addition of new neurons generated in the subventricular zone, were observed in the injured bulb. These data indicate that olfactory sensory neurons can adapt to reductions in their normal target field by obtaining sufficient support from remaining or alternative cell sources to survive and maintain their projections.

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In VitroGeneration of Adult Rat Olfactory Sensory Neurons and Regulation of Maturation by Coculture with CNS Tissues

Cited by 19 publications

References 44 publications

Maintaining epitheliopoietic potency when culturing olfactory progenitors

Maintaining epitheliopoietic potency when culturing olfactory progenitors

Interaction between astrocytes and adult subventricular zone precursors stimulates neurogenesis

Long‐term survival of olfactory sensory neurons after target depletion

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