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Spiral ganglion stem cells can be propagated and differentiated into neurons and glia.

Diensthuber M, Zecha V, Wagenblast J, Arnhold S, Edge AS, Stöver T - Biores Open Access (2014)

Bottom Line: Importantly, spiral ganglion sphere cells maintain their major stem cell characteristics after repeated propagation, which enables the culture of spheres for an extended period of time.In this work, we also demonstrate that differentiated sphere-derived cell populations not only adopt the immunophenotype of mature spiral ganglion cells but also develop distinct ultrastructural features of neurons and glial cells.Thus, our work provides further evidence that self-renewing spiral ganglion stem cells might serve as a promising source for the regeneration of lost auditory neurons.

View Article: PubMed Central - PubMed

Affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Frankfurt am Main , Goethe University, Frankfurt am Main, Germany . ; Department of Otology and Laryngology, Harvard Medical School , Boston, Massachusetts. ; Eaton-Peabody Laboratory, Massachusetts Eye and Ear Infirmary , Boston, Massachusetts.

ABSTRACT
The spiral ganglion is an essential functional component of the peripheral auditory system. Most types of hearing loss are associated with spiral ganglion cell degeneration which is irreversible due to the inner ear's lack of regenerative capacity. Recent studies revealed the existence of stem cells in the postnatal spiral ganglion, which gives rise to the hope that these cells might be useful for regenerative inner ear therapies. Here, we provide an in-depth analysis of sphere-forming stem cells isolated from the spiral ganglion of postnatal mice. We show that spiral ganglion spheres have characteristics similar to neurospheres isolated from the brain. Importantly, spiral ganglion sphere cells maintain their major stem cell characteristics after repeated propagation, which enables the culture of spheres for an extended period of time. In this work, we also demonstrate that differentiated sphere-derived cell populations not only adopt the immunophenotype of mature spiral ganglion cells but also develop distinct ultrastructural features of neurons and glial cells. Thus, our work provides further evidence that self-renewing spiral ganglion stem cells might serve as a promising source for the regeneration of lost auditory neurons.

No MeSH data available.


Related in: MedlinePlus

Differentiation of spiral ganglion-derived sphere populations into mature cells. (A) Light microscopic appearance of an adherently growing, differentiating spiral ganglion sphere. (B, C) After a 10-day differentiation period neuron-like cells expressing the neuronal marker MAP-2 can be found among plenty of GFAP-positive glia-like cells. When a BrdU-pulse is added to sphere cultures, nuclear BrdU incorporation can be found in neuron-like (D) and glia-like (E) cells differentiated from these spheres, indicating that spiral ganglion sphere-derived differentiated cells arise from dividing stem/progenitor cells. Scale bar=250 μm in A, 100 μm in B and E, and 120 μm in D.
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f8: Differentiation of spiral ganglion-derived sphere populations into mature cells. (A) Light microscopic appearance of an adherently growing, differentiating spiral ganglion sphere. (B, C) After a 10-day differentiation period neuron-like cells expressing the neuronal marker MAP-2 can be found among plenty of GFAP-positive glia-like cells. When a BrdU-pulse is added to sphere cultures, nuclear BrdU incorporation can be found in neuron-like (D) and glia-like (E) cells differentiated from these spheres, indicating that spiral ganglion sphere-derived differentiated cells arise from dividing stem/progenitor cells. Scale bar=250 μm in A, 100 μm in B and E, and 120 μm in D.

Mentions: It has been shown previously that spiral ganglion-derived spheres have the potential to differentiate into cells that express the neuronal markers TUJ and NF-M and the glial cell marker GFAP.8–10 These studies suggest that specific culture conditions can promote sphere differentiation and lead to the acquisition of a mature neuronal or glial immunophenotype. Our data corroborate these findings: when we transferred floating spheres to gelatin-coated plates with media containing BDNF and NT-3, spheres attached, flattened, and started to differentiate (Fig. 8A, Supplementary Movie S1). After a 10-day differentiation period we found 1.9±0.2% MAP-2-expressing cells and 13.1±4.2% GFAP-expressing cells in sphere-derived cell colonies (Fig. 8B, C). MAP-2 is a stringent marker of neurons in the central nervous system, but has also been shown to be expressed in the perikarya and neurites of spiral ganglion neurons.30 To find out whether the neurons and glial cells differentiated from spiral ganglion sphere cells that were newly created after culture, we added BrdU for 48 h during sphere generation. When we analyzed differentiated cell populations deriving from these spheres, we observed nuclear BrdU incorporation in MAP-2–expressing neuron-like cells (Fig. 8D) and GFAP-expressing glia-like cells (Fig. 8E). This finding indicates that neurons and glia cells developed from dividing progenitors in vitro.


Spiral ganglion stem cells can be propagated and differentiated into neurons and glia.

Diensthuber M, Zecha V, Wagenblast J, Arnhold S, Edge AS, Stöver T - Biores Open Access (2014)

Differentiation of spiral ganglion-derived sphere populations into mature cells. (A) Light microscopic appearance of an adherently growing, differentiating spiral ganglion sphere. (B, C) After a 10-day differentiation period neuron-like cells expressing the neuronal marker MAP-2 can be found among plenty of GFAP-positive glia-like cells. When a BrdU-pulse is added to sphere cultures, nuclear BrdU incorporation can be found in neuron-like (D) and glia-like (E) cells differentiated from these spheres, indicating that spiral ganglion sphere-derived differentiated cells arise from dividing stem/progenitor cells. Scale bar=250 μm in A, 100 μm in B and E, and 120 μm in D.
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Related In: Results  -  Collection

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f8: Differentiation of spiral ganglion-derived sphere populations into mature cells. (A) Light microscopic appearance of an adherently growing, differentiating spiral ganglion sphere. (B, C) After a 10-day differentiation period neuron-like cells expressing the neuronal marker MAP-2 can be found among plenty of GFAP-positive glia-like cells. When a BrdU-pulse is added to sphere cultures, nuclear BrdU incorporation can be found in neuron-like (D) and glia-like (E) cells differentiated from these spheres, indicating that spiral ganglion sphere-derived differentiated cells arise from dividing stem/progenitor cells. Scale bar=250 μm in A, 100 μm in B and E, and 120 μm in D.
Mentions: It has been shown previously that spiral ganglion-derived spheres have the potential to differentiate into cells that express the neuronal markers TUJ and NF-M and the glial cell marker GFAP.8–10 These studies suggest that specific culture conditions can promote sphere differentiation and lead to the acquisition of a mature neuronal or glial immunophenotype. Our data corroborate these findings: when we transferred floating spheres to gelatin-coated plates with media containing BDNF and NT-3, spheres attached, flattened, and started to differentiate (Fig. 8A, Supplementary Movie S1). After a 10-day differentiation period we found 1.9±0.2% MAP-2-expressing cells and 13.1±4.2% GFAP-expressing cells in sphere-derived cell colonies (Fig. 8B, C). MAP-2 is a stringent marker of neurons in the central nervous system, but has also been shown to be expressed in the perikarya and neurites of spiral ganglion neurons.30 To find out whether the neurons and glial cells differentiated from spiral ganglion sphere cells that were newly created after culture, we added BrdU for 48 h during sphere generation. When we analyzed differentiated cell populations deriving from these spheres, we observed nuclear BrdU incorporation in MAP-2–expressing neuron-like cells (Fig. 8D) and GFAP-expressing glia-like cells (Fig. 8E). This finding indicates that neurons and glia cells developed from dividing progenitors in vitro.

Bottom Line: Importantly, spiral ganglion sphere cells maintain their major stem cell characteristics after repeated propagation, which enables the culture of spheres for an extended period of time.In this work, we also demonstrate that differentiated sphere-derived cell populations not only adopt the immunophenotype of mature spiral ganglion cells but also develop distinct ultrastructural features of neurons and glial cells.Thus, our work provides further evidence that self-renewing spiral ganglion stem cells might serve as a promising source for the regeneration of lost auditory neurons.

View Article: PubMed Central - PubMed

Affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Frankfurt am Main , Goethe University, Frankfurt am Main, Germany . ; Department of Otology and Laryngology, Harvard Medical School , Boston, Massachusetts. ; Eaton-Peabody Laboratory, Massachusetts Eye and Ear Infirmary , Boston, Massachusetts.

ABSTRACT
The spiral ganglion is an essential functional component of the peripheral auditory system. Most types of hearing loss are associated with spiral ganglion cell degeneration which is irreversible due to the inner ear's lack of regenerative capacity. Recent studies revealed the existence of stem cells in the postnatal spiral ganglion, which gives rise to the hope that these cells might be useful for regenerative inner ear therapies. Here, we provide an in-depth analysis of sphere-forming stem cells isolated from the spiral ganglion of postnatal mice. We show that spiral ganglion spheres have characteristics similar to neurospheres isolated from the brain. Importantly, spiral ganglion sphere cells maintain their major stem cell characteristics after repeated propagation, which enables the culture of spheres for an extended period of time. In this work, we also demonstrate that differentiated sphere-derived cell populations not only adopt the immunophenotype of mature spiral ganglion cells but also develop distinct ultrastructural features of neurons and glial cells. Thus, our work provides further evidence that self-renewing spiral ganglion stem cells might serve as a promising source for the regeneration of lost auditory neurons.

No MeSH data available.


Related in: MedlinePlus