论文部分内容阅读
目的研究成人和成年狗嗅黏膜成鞘细胞(OECs)的分离与培养的方法,为探索在人体和大型动物模型的自体嗅黏膜OECs移植修复脊髓损伤奠定基础。方法采用差速贴壁方法分别从成人和成年比格狗的嗅黏膜分离出OECs,分别培养25 d,在第6 d1、0 d和25 d进行形态学观察,最后进行OECs特异标记物NGFRp75的免疫细胞化学染色,鉴定成鞘细胞。结果原代培养的成人和成年狗嗅黏膜OECs在培养10 d后明显增多,25 d的OECs成熟,具有很长的突起,其形态多为双极和三极,NGFRp75的免疫组织化学染色呈阳性。本实验条件下,狗嗅黏膜OECs的生长状态比成人的好。而未进行差速贴壁的培养细胞则几乎均呈成纤维样细胞。结论差速贴壁的方法可以分离出较为纯化的成人、成年狗鼻腔嗅黏膜OECs。
Objective To study the isolation and culture of olfactory ensheathing cells (OECs) from adult and adult dogs, and to lay the foundation for the exploration of repairing spinal cord injury by autologous olfactory mucosa OECs transplantation in human and large animal models. Methods OECs were isolated from the olfactory mucosa of adult and adult beagle dogs by differential adherence method. The cultured animals were cultured for 25 days, respectively. Morphological observation was performed on the 6th day, the 1st day and the 25th day. Finally, the OECs specific markers NGFRp75 Immunocytochemical staining, identification of sheath cells. Results OECs of primary cultured adult and adult dogs were significantly increased after 10 days of culture. OECs of 25 days old were mature with long protuberances, most of them were bipolar and tripolar, and immunohistochemical staining of NGFRp75 was positive . Under the experimental conditions, the growth of OECs in dog olfactory mucosa is better than that in adults. The cultured cells without differential adherence almost showed fibroblast-like cells. Conclusion Differential adherence method can isolate more purified nasal olfactory mucosa OECs from adult and adult dogs.