涎腺腺样囊性癌细胞与鸡胚背根神经节共培养的实验研究
Experimental study on co-culture of salivary adenoid cystic carcinoma cells and ganglia
摘要目的 观察涎腺腺样囊性癌(salivary adenoid cystic carcinoma,SACC) 83细胞与鸡胚背根神经节(dorsal root ganglia,DRG)共培养模型中肿瘤细胞对神经节突起生长的诱向作用,并初步探讨发挥诱导作用的因子.方法采用玻璃底培养皿构建SACC与DRG共培养模型,SACC-83细胞接种至中央孔内,DRG组织环绕接种1周.观察神经节突起的生长情况,再分别以不同培养液对DRG培养,实验分6组:1组每孔中各加入SACC-83条件培养基、2组加入鼠成骨细胞系MC3T3-E1条件培养基、3组加入牙龈成纤维细胞条件培养基、4组加入无血清高糖达尔伯克改良伊格尔培养基(Dulbecco's modification of Eagle's medium with high glucose,H-DMEM)、5组加入含15%胎牛血清的H-DMEM、6组加入SACC-83细胞裂解液对DRG培养36 h,观察神经节突起的生长情况.结果培养36 h玻璃底培养皿共培养模型中多数神经节突起呈向中心趋向生长的状态;1组有明显的促神经节突起生长的作用,4~6组无促神经节突起生长的作用.结论以玻璃底培养皿构建肿瘤-神经共培养模型能较好地观察肿瘤细胞构成的微环境对神经突起生长的趋化作用,该作用与细胞分泌的某些特异性神经活性物质有关,该现象的存在可能与SACC嗜神经侵袭特性有关.
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abstractsObjective To construct the co-culture models of salivarya denoid cystic carcinoma (SACC) cells and dorsal root ganglia (DRG) of chickens and investigate the promotive effects of SACC on neural tissue.Methods Glass-base culture dish was adopted to construct co-culture model of SACC-83 cells and DRG. SACC-83 cells were seeded in the medium pore with DRG around them. Outgrowth of neuronal processes was observed.Then DRG was cultured in the conditioned medium of SACC-83,with the groups of conditioned medium of MC3T3-E1 and HGF,the group of cell lysis buffer,the groups of serum free medium and serum-plus medium as the controls.Outgrowth of neuronal processes was also recorded and compared with control groups.Results In the co-culture model of tumor and neuronal tissue,SACC-83cells produced a suitable microenvironment in which neuronal processes remarkably grow. Neuronal processes of most DRG displayed growth tendency toward SACC.The group of conditioned medium from SACC-83 manifested obvious promotive effects on DRG. Conclusions Co-culture model of tumor and neuronal tissue was successfully constructed,with which the promotive effects of tumor on outgrowth of neuronal processes could be observed.So hypothesized that SACC could secrete some neurotrophic factors to guide peripheral nerves gemmating and to trigger the cascade of the neural invasion in succession.
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