嗅上皮的组织块培养法

目的建立并优化组织块法嗅觉受体神经元（olfactory receptor neurons,ORNs）原代培养体系。方法以鼠尾胶原为培养底物,取新生小鼠鼻中隔和筛甲的嗅上皮进行原代培养。观察包括ORNs在内的各种细胞的形态和分布特点,经免疫组化和扫描电镜证实本原代培养体系内各种细胞的属性以及ORNs的分化程度。结果组织块培养法所得为一混杂的培养体系,除ORNs外还有基底细胞、嗅鞘细胞、纺锤形细胞及成纤维细胞等。其中基底细胞和纺锤形细胞在形态和免疫表型上均具有ORNs前体细胞的特点。优化培养体系后,可得到分化成熟的ORNs并可在体外稳定存活9天以上。结论本培养方法避免了使用消化酶,可以稳定的得到分化成熟的ORNs。

Explant culture of olfactory epithelium

OBJECTIVE The aim of the study was to setup and optimize the explant culture system of olfactory receptor neurons（ORNs）.METHODS Rat tail collagen was used as the culture substratum.The olfactory epithelium（OE）from the septum and ethmoid turbinates of newborn mice was harvested,plated and cultured.The morphology and growth pattern of cultured cells including ORNs were studied under inverted microscope and scanning electron microscope.The phenotype and maturity of the cultured cells were determined by means of Immunocytochemistry.RESULTS After optimization of the culture condition,fully developed ORNs were achieved.The OE explant culture was a heterogeneous culture system,which consisted of basal cells,olfactory ensheathing cells（OECs）,fusiform cells,and fibroblasts besides ORNs.The basal cells and fusiform cells were characterized as precursors of ORNs.The ORNs maintained stably beyond 9 days in vitro.CONCLUSION Using explant culture,which devoid the potential damage of any enzyme,fully developed ORNs could be stably achieved.