Wang Han, Deng Liling, Qin Xuanhe
School of Life Science and Technology, Tongji University, Shanghai 200092, China.
Shanghai East Hospital, Tongji University, Shanghai 200120, China.
Sheng Wu Gong Cheng Xue Bao. 2023 Jan 25;39(1):318-336. doi: 10.13345/j.cjb.220244.
Olfactory epithelium, which detects and transmits odor signals, is critical for the function of olfactory system. Olfactory epithelium is able to recover spontaneously after injury under normal circumstances, but this ability is dampened in certain diseases or senility, which causes olfactory dysfunction. The olfactory epithelium consists of basal cells, sustentacular cells and olfactory sensory neurons. In order to develop an olfactory epithelial organoid containing multiple olfactory cell types , we used three-dimensional culture model and small molecules screening. This organoid system consists of horizontal basal-like cells, globose basal-like cells, sustentacular-like cells and olfactory sensory neurons-like cells. Through statistical analysis of clone diameter, immunofluorescence staining and qPCR detection of the expression level of related marker genes. We identified a series of growth factors and small molecule compounds that affected the proliferation, composition and gene expression of the organoids. CHIR-99021, an activator of Wnt signaling pathway, increased the colony formation and proliferation rate of olfactory epithelial organoids and the expression level of marker genes of olfactory sensory neurons-like cells. In addition, each factor in the culture system increased the proportion of c-Kit-positive globose basal-like cell colonies in organoids. Moreover, EGF and vitamin C were both beneficial to the expression of horizontal basal-like cell marker genes in organoids. The established olfactory epithelial organoid system mimicked the process of olfactory epithelial stem cells differentiating into various olfactory epithelial cell types, thus providing a research model for studying olfactory epithelial tissue regeneration, the pathological mechanism of olfactory dysfunction and drug screening for olfactory dysfunction treatment.
嗅觉上皮能够检测并传递气味信号，对嗅觉系统的功能至关重要。在正常情况下，嗅觉上皮损伤后能够自发恢复，但在某些疾病或衰老过程中，这种能力会受到抑制，从而导致嗅觉功能障碍。嗅觉上皮由基底细胞、支持细胞和嗅觉感觉神经元组成。为了构建包含多种嗅觉细胞类型的嗅觉上皮类器官，我们采用了三维培养模型和小分子筛选技术。该类器官系统由水平基底样细胞、球状基底样细胞、支持样细胞和嗅觉感觉神经元样细胞组成。通过对克隆直径进行统计分析、免疫荧光染色以及对相关标记基因表达水平进行qPCR检测，我们鉴定出了一系列影响类器官增殖、组成和基因表达的生长因子和小分子化合物。CHIR-99021是一种Wnt信号通路激活剂，它提高了嗅觉上皮类器官的集落形成和增殖率以及嗅觉感觉神经元样细胞标记基因的表达水平。此外，培养体系中的每种因子都增加了类器官中c-Kit阳性球状基底样细胞集落的比例。而且，表皮生长因子（EGF）和维生素C都有利于类器官中水平基底样细胞标记基因的表达。所建立的嗅觉上皮类器官系统模拟了嗅觉上皮干细胞分化为各种嗅觉上皮细胞类型的过程，从而为研究嗅觉上皮组织再生、嗅觉功能障碍的病理机制以及嗅觉功能障碍治疗的药物筛选提供了一个研究模型。
