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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

Spiral ganglion-derived spheres are floating colonies with high proliferative capacity. Spheres that form in culture media containing BrdU show incorporation of the thymidine analog in a significant proportion of the sphere cells which can be observed in primary spheres (A). Following propagation, BrdU is also incorporated during the formation of secondary (B) and tertiary spheres (C). Nuclear expression of Ki-67, a marker associated with the active phases of the cell cycle, indicates the proliferative activity of primary spheres (D) and subsequent sphere generations (E, F). (G, H) Quantification of BrdU and Ki-67 expression in the primary, secondary, and tertiary generation of spiral ganglion-derived spheres. Data are the mean±SD from three independent experiments. Scale bar=100 μm.
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f6: Spiral ganglion-derived spheres are floating colonies with high proliferative capacity. Spheres that form in culture media containing BrdU show incorporation of the thymidine analog in a significant proportion of the sphere cells which can be observed in primary spheres (A). Following propagation, BrdU is also incorporated during the formation of secondary (B) and tertiary spheres (C). Nuclear expression of Ki-67, a marker associated with the active phases of the cell cycle, indicates the proliferative activity of primary spheres (D) and subsequent sphere generations (E, F). (G, H) Quantification of BrdU and Ki-67 expression in the primary, secondary, and tertiary generation of spiral ganglion-derived spheres. Data are the mean±SD from three independent experiments. Scale bar=100 μm.

Mentions: Spheres isolated from the central nervous system or the inner ear arise from proliferating cells and sphere cells express various neural stem/progenitor cell markers.8,9,18,21,22,29 We quantitatively analyzed and compared proliferation activity and marker expression in primary spheres and spheres obtained by repeated propagation (secondary and tertiary spheres). We found robust BrdU incorporation in the nuclei of primary sphere cells (75.7±0.8%) indicating that these cells underwent cell division during sphere formation in the neurosphere assay (Fig. 6A,G). The detection of Ki-67 expression, a marker for cell proliferation, in 4.7±0.4% of the primary sphere cells is a further indication of their proliferative activity (Fig. 6D,H). A significant number of primary sphere cells expressed the neural stem cell marker nestin (85.8±3.7%) (Fig. 7A,B,E). To find out whether spheres maintain their stem/progenitor cell status and proliferative activity after propagation, we analyzed the secondary and tertiary sphere generation. Secondary and tertiary spheres also showed significant proliferative activity as indicated by BrdU incorporation (38.2±1.6% and 10.4±4.3% of the sphere cells) (Fig. 6B,C,G) and Ki-67 expression (3.1±0.2% and 2.6±0.7% of the sphere cells) (Fig. 6E,F,H). Likewise, propagated spheres seem to keep their stem/progenitor cell status since we detected robust expression of nestin in the sphere cells (54.3±2.2% of the secondary sphere cells, 42.0±6.1% of the tertiary sphere cells) (Fig. 7C–E).


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)

Spiral ganglion-derived spheres are floating colonies with high proliferative capacity. Spheres that form in culture media containing BrdU show incorporation of the thymidine analog in a significant proportion of the sphere cells which can be observed in primary spheres (A). Following propagation, BrdU is also incorporated during the formation of secondary (B) and tertiary spheres (C). Nuclear expression of Ki-67, a marker associated with the active phases of the cell cycle, indicates the proliferative activity of primary spheres (D) and subsequent sphere generations (E, F). (G, H) Quantification of BrdU and Ki-67 expression in the primary, secondary, and tertiary generation of spiral ganglion-derived spheres. Data are the mean±SD from three independent experiments. Scale bar=100 μm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4048968&req=5

f6: Spiral ganglion-derived spheres are floating colonies with high proliferative capacity. Spheres that form in culture media containing BrdU show incorporation of the thymidine analog in a significant proportion of the sphere cells which can be observed in primary spheres (A). Following propagation, BrdU is also incorporated during the formation of secondary (B) and tertiary spheres (C). Nuclear expression of Ki-67, a marker associated with the active phases of the cell cycle, indicates the proliferative activity of primary spheres (D) and subsequent sphere generations (E, F). (G, H) Quantification of BrdU and Ki-67 expression in the primary, secondary, and tertiary generation of spiral ganglion-derived spheres. Data are the mean±SD from three independent experiments. Scale bar=100 μm.
Mentions: Spheres isolated from the central nervous system or the inner ear arise from proliferating cells and sphere cells express various neural stem/progenitor cell markers.8,9,18,21,22,29 We quantitatively analyzed and compared proliferation activity and marker expression in primary spheres and spheres obtained by repeated propagation (secondary and tertiary spheres). We found robust BrdU incorporation in the nuclei of primary sphere cells (75.7±0.8%) indicating that these cells underwent cell division during sphere formation in the neurosphere assay (Fig. 6A,G). The detection of Ki-67 expression, a marker for cell proliferation, in 4.7±0.4% of the primary sphere cells is a further indication of their proliferative activity (Fig. 6D,H). A significant number of primary sphere cells expressed the neural stem cell marker nestin (85.8±3.7%) (Fig. 7A,B,E). To find out whether spheres maintain their stem/progenitor cell status and proliferative activity after propagation, we analyzed the secondary and tertiary sphere generation. Secondary and tertiary spheres also showed significant proliferative activity as indicated by BrdU incorporation (38.2±1.6% and 10.4±4.3% of the sphere cells) (Fig. 6B,C,G) and Ki-67 expression (3.1±0.2% and 2.6±0.7% of the sphere cells) (Fig. 6E,F,H). Likewise, propagated spheres seem to keep their stem/progenitor cell status since we detected robust expression of nestin in the sphere cells (54.3±2.2% of the secondary sphere cells, 42.0±6.1% of the tertiary sphere cells) (Fig. 7C–E).

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