Journal
ACTA OTO-LARYNGOLOGICA
Volume 132, Issue 7, Pages 693-701Publisher
INFORMA HEALTHCARE
DOI: 10.3109/00016489.2012.657358
Keywords
Sphere; auditory pathway; BrdU; microarray
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Funding
- Kansai Medical University
- 'Academic Frontier' Project for Private Universities
- MEXT (Ministry of Education, Culture, Sports, Science and Technology)
- Japanese Ministry of Education, Culture, Sports, Science and Technology [16791029, 18790740]
- Society for Research Aid in Otolaryngology, Kansai Medical University
- Grants-in-Aid for Scientific Research [16791029, 18790740] Funding Source: KAKEN
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Conclusion: We characterized side population (SP) cells in the cochlear nucleus (CN). Some genes of stem/progenitor markers in sorted SP cells were identified by microarray analysis and RT-PCR. Furthermore, some cells in the CN also demonstrated self-renewal and clonal expansion activities. These results suggest that tissue stem/progenitor like cells would be identified and characterized as a slow cycling and immaturity in SP cells of CN. Objectives: SP cells were sorted and characterized as regards their activity in the CN in order to identify the tissue progenitor/stem cells in the auditory nervous system. Methods: Bromodeoxyuridine (BrdU)-injected mice were prepared and the long-term BrdU-retaining cells were detected by flow cytometry. Gene expression of SP and MP cells was analyzed by microarray analysis and RT-PCR. SP cells were cultured in conditioned medium to expand stem/progenitor cells in vitro and to estimate the spheroid-forming activity of stem cells. Results: In all, 1% of cells in the CN were detected as BrdU-positive. SP cells were detected at a frequency of 4.4% and expressed stem/progenitor markers, Abcb1b, Abcg2, Sca1, Notch1, Notch4, Hes1, and Jag1 in microarray analysis. Expression of Abcb1b, Abcg2, Sca1, Oct3/4, and Sox2 as determined by RT-PCR was supported by the microarray data. CN cells also had sphere-forming activity in young mice, but this activity was decreased by aging.
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