Expansion of murine and human olfactory epithelium/mucosa colonies and generation of mature olfactory sensory neurons under chemically defined conditions.
Expansion of murine and human olfactory epithelium/mucosa colonies and generation of mature olfactory sensory neurons under chemically defined conditions.
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在化学定义的条件下扩增小鼠和人类嗅觉上皮/粘膜集落并生成成熟的嗅觉感觉神经元
作者：
Ren W;Wang L;Zhang X;Feng X;Zhuang L;Jiang N;Xu R;Li X;Wang P;Sun X;Yu H;Yu Y
嗅觉功能障碍，包括嗅觉减退和嗅觉丧失，影响着全世界约1亿人，其根本原因尚未完全了解。老年人和嗅觉障碍患者存在嗅感觉神经元变性和球状基底细胞不能产生嗅感觉神经元的现象。因此，嗅觉干细胞可能作为一种有前途的工具，以取代失活的球状基底细胞和感觉神经元。研究方法：我们建立了小鼠嗅上皮细胞的克隆扩增以及使用基于Matrigel的三维系统从人嗅粘膜的集落生长。这些殖民地的特点是对嗅上皮细胞标志物的免疫染色和钙成像的反应气味。化学添加进行了优化，以促进Lgr 5的表达，集落生长和感觉神经元的产生，测试通过定量PCR和免疫染色对祖细胞和神经元标记。利用RNA-Seq技术分析了不同培养基和化学混合物条件下菌落间多种信号通路的差异转录组。结果如下：在确定的培养基中，我们发现VPA和CHIR 99021诱导最高的Lgr 5表达水平，而LY 411575导致小鼠集落中OMP+成熟感觉神经元的最丰富产量。不同的基础培养基与药物鸡尾酒导致明显的形态学改变，从填充到囊性外观，伴随着大量的转录变化，在多个信号转导通路。人集落中感觉神经元的产生通过TGF-β信号传导受到影响，而Lgr 5表达和细胞增殖受VPA调节。结论：我们的研究结果表明，体外靶向扩增嗅上皮/粘膜集落可能会发现新的来源，以细胞增殖为基础的治疗嗅觉丧失。
Olfactory dysfunctions, including hyposmia and anosmia, affect ~100 million people around the world and the underlying causes are not fully understood. Degeneration of olfactory sensory neurons and incapacity of globose basal cells to generate olfactory sensory neurons are found in elder people and patients with smell disorders. Thus, olfactory stem cell may function as a promising tool to replace inactivated globose basal cells and to generate sensory neurons. Methods: We established clonal expansion of cells from the murine olfactory epithelium as well as colony growth from human olfactory mucosa using Matrigel-based three-dimensional system. These colonies were characterized by immunostaining against olfactory epithelium cellular markers and by calcium imaging of responses to odors. Chemical addition was optimized to promote Lgr5 expression, colony growth and sensory neuron generation, tested by quantitative PCR and immunostaining against progenitor and neuronal markers. The differential transcriptomes in multiple signaling pathways between colonies under different base media and chemical cocktails were determined by RNA-Seq. Results: In defined culture media, we found that VPA and CHIR99021 induced the highest Lgr5 expression level, while LY411575 resulted in the most abundant yield of OMP+ mature sensory neurons in murine colonies. Different base culture media with drug cocktails led to apparent morphological alteration from filled to cystic appearance, accompanied with massive transcriptional changes in multiple signaling pathways. Generation of sensory neurons in human colonies was affected through TGF-β signaling, while Lgr5 expression and cell proliferation was regulated by VPA. Conclusion: Our findings suggest that targeting expansion of olfactory epithelium/mucosa colonies in vitro potentially results in discovery of new source to cell replacement-based therapy against smell loss.
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影响因子：
9
作者：
Fan JR;Lee HT;Lee W;Lin CH;Hsu CY;Hsieh CH;Shyu WC
通讯作者：
Shyu WC
影响因子：
2.9
作者：
Baud, Olivia;Etter, Sylvain;Pick, Horst
通讯作者：
Pick, Horst
影响因子：
5.3
作者：
Child, Kevin M.;Herrick, Daniel B.;Jang, Woochan
通讯作者：
Jang, Woochan
DOI：
10.1111/j.1749-6632.1998.tb10650.x
发表时间：
1998-01-01
期刊：
OLFACTION AND TASTE XII
影响因子：
--
作者：
Hoffman, HJ;Ishii, EK;Macturk, RH
通讯作者：
Macturk, RH
影响因子：
3.7
作者：
Abranches E;Silva M;Pradier L;Schulz H;Hummel O;Henrique D;Bekman E
通讯作者：
Bekman E