Ooka Hisashi, Kanda Seiji, Okazaki Haruka, Suzuki Hiroko, Mishima Kenji, Saito Ichiro, Yagi Masao, Tomoda Koichi, Nishiyama Toshimasa
Regeneration Research Center for Intractable Diseases, Kansai Medical University, Osaka, Japan.
Acta Otolaryngol. 2012 Jul;132(7):693-701. doi: 10.3109/00016489.2012.657358. Epub 2012 Jun 5.
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.
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.
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.
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.
我们对蜗神经核（CN）中的侧群（SP）细胞进行了表征。通过微阵列分析和逆转录聚合酶链反应（RT-PCR）鉴定了分选的SP细胞中一些干/祖细胞标志物的基因。此外，CN中的一些细胞还表现出自我更新和克隆扩增活性。这些结果表明，组织干/祖细胞样细胞可在CN的SP细胞中被鉴定为慢循环和未成熟细胞并进行表征。
对SP细胞进行分选并表征其在CN中的活性，以鉴定听觉神经系统中的组织祖细胞/干细胞。
制备注射了溴脱氧尿苷（BrdU）的小鼠，并通过流式细胞术检测长期保留BrdU的细胞。通过微阵列分析和RT-PCR分析SP和MP细胞的基因表达。将SP细胞在条件培养基中培养以体外扩增干/祖细胞，并评估干细胞的球形成活性。
总共检测到CN中1%的细胞为BrdU阳性。SP细胞的检测频率为4.4%，在微阵列分析中表达干/祖细胞标志物Abcb1b、Abcg2、Sca1、Notch1、Notch4、Hes1和Jag1。微阵列数据支持RT-PCR测定的Abcb1b、Abcg2、Sca1、Oct3/4和Sox2的表达。CN细胞在幼鼠中也具有球形成活性，但这种活性会随着年龄增长而降低。
