Ito Daisuke, Ibanez Chrystelle, Ogawa Hiroyuki, Franklin Robin J M, Jeffery Nick D
Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UK.
Am J Vet Res. 2006 Jun;67(6):1050-6. doi: 10.2460/ajvr.67.6.1050.
To evaluate the numbers and proportions of olfactory ensheathing cells (OECs) in cell cultures derived from the olfactory bulb (OB) and olfactory mucosa of dogs.
7 dogs.
OB tissue and olfactory mucosa from the nasal cavity and frontal sinus were obtained from euthanatized dogs and prepared for cell culture. At 7, 14, and 21 days of culture in vitro, numbers and proportions of OECs, astrocytes, and fibroblasts were determined via immunocytochemistry. Antibody against the low-affinity nerve growth factor receptor p75 was used to identify OECs, antibody against glial fibrillary acidic protein was used to identify astrocytes, and antibody against fibronectin was used to identify fibroblasts.
Cultured OECs derived from the olfactory mucosa of the nasal cavity and frontal sinus had similar characteristics. However, whereas OECs in the OB cell cultures constituted approximately 50% of the cells at 7 days and approximately 75% at 21 days the proportion of OECs in cultures derived from both mucosal types was much lower, with approximately 40% OECs at 7 days and approximately 25% at 21 days. Analysis of OEC numbers revealed that these changes were accompanied by corresponding decreases and increases in the population of cells with fibronectin receptors.
Although olfactory mucosal cell cultures yielded a sufficient number of OECs for spinal cord transplantation procedures in dogs, modification of culture conditions would be required to ensure that the derived cell population contained a sufficient proportion of OECs.
评估源自犬嗅球(OB)和嗅黏膜的细胞培养物中嗅鞘细胞(OECs)的数量和比例。
7只犬。
从安乐死的犬获取OB组织以及鼻腔和额窦的嗅黏膜,并制备用于细胞培养。在体外培养7、14和21天时,通过免疫细胞化学测定OECs、星形胶质细胞和成纤维细胞的数量和比例。使用抗低亲和力神经生长因子受体p75的抗体鉴定OECs,使用抗胶质纤维酸性蛋白的抗体鉴定星形胶质细胞,使用抗纤连蛋白的抗体鉴定成纤维细胞。
源自鼻腔和额窦嗅黏膜的培养OECs具有相似特征。然而,OB细胞培养物中的OECs在7天时约占细胞总数的50%,在21天时约占75%,而源自两种黏膜类型的培养物中OECs的比例要低得多,7天时约为40%,21天时约为25%。对OEC数量的分析表明,这些变化伴随着具有纤连蛋白受体的细胞群体相应的减少和增加。
尽管嗅黏膜细胞培养物产生了足够数量的OECs用于犬脊髓移植手术,但需要调整培养条件以确保所获得的细胞群体中含有足够比例的OECs。