Comparing the cultivated cochlear cells derived from neonatal and adult mouse

Journal of Translational Medicine

Yanzhong Zhang²  Masaaki Tokuda¹  Liangliang Yang²  Xianliu Wang²  Jing Xie²  Youyi Dong¹  Xiangxin Lou² 

[1]Department of Cell Physiology, Faculty of Medicine, Kagawa University, 1750-1 Ikenobe, Miki-cho, Kita-gun, Kagawa 761-0793, Japan

[2]Department of Bioengineering, College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, 2999 North Renmin Road, Shanghai 201620, China

关键词: Progenitor cell;    Sphere;    Hair cell;    Cochlea;   

Others  :  810852
DOI  :  10.1186/1479-5876-12-150

 received in 2013-11-21, accepted in 2014-05-21,  发布年份 2014

【 摘 要 】

Background

Previous reports showed the presence of limited numbers of stem cells in neonatal murine cochlear sensory epithelia and these cells are progressively lost during the postnatal development. The goal of this study was to investigate whether stem cells can be derived from mature mouse cochleae under suspension culture conditions, and to analyze the expression of the stem cell and inner ear progenitor cell markers in cells dissociated from neonatal and adult mouse organs of Corti.

Methods

Organs of Corti were dissected from postnatal day 1 (P1) or postnatal day 60 (P60) mouse. The dissociated cells were cultivated under suspension cultures conditions. Reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry were conducted for phenotype characterization.

Results

The number of cochlear stem cells (otospheres) yielded from P1 organ of Corti was significantly higher than that of the P60 organ of Corti. RT-PCR analyses showed that the stem markers, such as nanog, sox2, klf4, and nestin can be found to be distributed similarly in the cells derived from both of organisms, but the inner ear developmental/progenitor cell markers showed lower expression in P60 organ of Corti compared to P1. Immunocytochemistry results also revealed the evidence that P60 otospheres lacking of differentiation potential in vitro, which opposed to the strong differentiation potential of otospheres at P1 stage.

Conclusions

Our findings suggest that the loss of numbers and features of stem cells in the adult organ of Corti is associated with the substantial down-regulation of inner ear progenitor key-markers during maturation of the cells in organ of Corti.

【 授权许可】

   
2014 Lou et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Bekman EP, Costa A, Cristovao AM, Henrique D: Stem cell therapy for inner ear hair cell regeneration. Int J Dev Neurosci 2010, 28:667-668.
• [2]Beisel K, Hansen L, Soukup G, Fritzsch B: Regenerating cochlear hair cells: quo vadis stem cell. Cell Tissue Res 2008, 333:373-379.
• [3]Chao T-T, Wang C-H, Chen H-C, Shih C-P, Sytwu H-K, Huang K-L, Chen S-Y: Adherent culture conditions enrich the side population obtained from the cochlear modiolus-derived stem/progenitor cells. Int J Pediatr Otorhinolaryngol 2013, 77:779-784.
• [4]Diensthuber M, Oshima K, Heller S: Stem/Progenitor Cells Derived from the Cochlear Sensory Epithelium Give Rise to Spheres with Distinct Morphologies and Features. J Assoc Res Otolaryngol 2009, 10:173-190.
• [5]Oshima K, Grimm CM, Corrales CE, Senn P, Moneder RM, Geleoc GSG, Edge A, Holt JR, Heller S: Differential distribution of stem cells in the auditory and vestibular organs of the inner ear. J Assoc Res Otolaryngol 2007, 8:18-31.
• [6]Jongkamonwiwat N, Zine A, Rivolta MN: Stem cell based therapy in the inner ear: appropriate donor cell types and routes for transplantation. Curr Drug Targets 2010, 11:888-897.
• [7]Malgrange B, Belachew S, Thiry M, Nguyen L, Rogister B, Alvarez ML, Rigo JM, Van de Water TR, Moonen G, Lefebvre PP: Proliferative generation of mammalian auditory hair cells in culture. Mech Dev 2002, 112:79-88.
• [8]Wang ZM, Jiang HQ, Yan YB, Wang YC, Shen YZ, Li W, Li HW: Characterization of proliferating cells from newborn mouse cochleae. Neuroreport 2006, 17:767-771.
• [9]Savary E, Sabourin JC, Santo J, Hugnot JP, Chabbert C, Van De Water T, Uziel A, Zine A: Cochlear stem/progenitor cells from a postnatal cochlea respond to Jagged1 and demonstrate that notch signaling promotes sphere formation and sensory potential. Mech Dev 2008, 125:674-686.
• [10]Senn P, Oshima K, Teo D, Grimm C, Heller S: Robust postmortem survival of murine vestibular and cochlear stem cells. J Assoc Res Otolaryngol 2007, 8:194-204.
• [11]Yerukhimovich MV, Bai L, Chen DHC, Miller RH, Alagramam KN: Identification and characterization of mouse cochlear stem cells. Dev Neurosci 2007, 29:251-260.
• [12]Zhai SQ, Shi L, Wang BE, Zheng GL, Song W, Hu YY, Gao WQ: Isolation and culture of hair cell progenitors from postnatal rat cochleae. J Neurobiol 2005, 65:282-293.
• [13]Zhang Y, Zhai S-Q, Shou J, Song W, Sun J-H, Guo W, Zheng G-l, Hu Y-Y, Gao W-Q: Isolation, growth and differentiation of hair cell progenitors from the newborn rat cochlear greater epithelial ridgele. J Neurosci Methods 2007, 164:271-279.
• [14]Oiticica J, Barboza-Junior LCM, Batissoco AC, Lezirovitz K, Mingroni-Netto RC, Haddad LA, Bento RF: Retention of progenitor cell phenotype in otospheres from guinea pig and mouse cochlea. J Transl Med 2010, 8:119. BioMed Central Full Text
• [15]Oshima K, Senn P, Heller S: Isolation of sphere-forming stem cells from the mouse inner ear. Methods Mol Biol 2009, 493:141-162.
• [16]Groves AK: The challenge of hair cell regeneration. Exp Biol Med 2010, 235:434-446.
• [17]Lin J, Feng L, Fukudome S, Hamajima Y, Huang T, Levine S: Cochlear stem cells/progenitors and degenerative hearing disorders. Curr Med Chem 2007, 14:2937-2943.
• [18]Lou X-X, Nakagawa T, Ohnishi H, Nishimura K, Ito J: Otospheres derived from neonatal mouse cochleae retain the progenitor cell phenotype after ex vivo expansions. Neurosci Lett 2013, 534:18-23.
• [19]Li HW, Liu H, Heller S: Pluripotent stem cells from the adult mouse inner ear. Nat Med 2003, 9:1293-1299.
• [20]Carricondo F, Cruz Iglesias M, Rodriguez F, Poch-Broto J, Gil-Loyzaga P: In vitro long-term development of cultured inner ear stem cells of newborn rat. Cell Tissue Res 2010, 342:13-19.
• [21]Edge A: Stem cells and regeneration of the inner ear. Neurosci Res 2008, 61:S25-S25.
• [22]El-Amraoui A, Petit C: Stem cell therapy in the inner ear: recent achievements and prospects. Med Sci 2010, 26:981-985.
• [23]Martinez-Moneder R, Edge ASB: Stem cells for the replacement of inner ear neurons and hair cells. Int J Dev Biol 2007, 51:655-661.
• [24]Senn P, Heller S: Stem-cell-based approaches for treating inner ear diseases. HNO 2008, 56:21-26.
• [25]Chen H-C, Sytwu H-K, Chang J-L, Wang H-W, Chen H-K, Kang B-H, Liu D-W, Chen C-H, Chao T-T, Wang C-H: Hypoxia enhances the stemness markers of cochlear stem/progenitor cells and expands sphere formation through activation of hypoxia-inducible factor-1alpha. Hear Res 2011, 275:43-52.
• [26]Kane KL, Longo-Guess CM, Gagnon LH, Ding D, Salvi RJ, Johnson KR: Genetic background effects on age-related hearing loss associated with Cdh23 variants in mice. Hear Res 2012, 283:80-88.
• [27]Yamamoto N, Tanigaki K, Tsuji M, Yabe D, Ito J, Honjo T: Inhibition of Notch/RBP-J signaling induces hair cell formation in neonate mouse cochleas. J Mol Med 2006, 84:37-45.
• [28]Lustig LR, Akil O: Cochlear gene therapy. Curr Opin Neurol 2012, 25:57-60.
• [29]Takahashi K, Yamanaka S: Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006, 126:663-676.
• [30]Polo JM, Anderssen E, Walsh RM, Schwarz BA, Nefzger CM, Lim SM, Borkent M, Apostolou E, Alaei S, Cloutier J, Bar-Nur O, Cheloufi S, Stadtfeld M, Figueroa ME, Robinton D, Natesan S, Melnick A, Zhu J, Ramaswamy S, Hochedlinger K: A molecular roadmap of reprogramming somatic cells into iPS cells. Cell 2012, 151:1617-1632.