人恶性脑膜瘤原代细胞的培养及小鼠颅内原位成瘤模型的建立

Primary culture of human malignant meningioma cells and its intracranial orthotopic transplantation in nude mice

目的培养稳定的恶性脑膜瘤原代细胞以及建立颅内原位成瘤模型。方法改良法培养术前高度可疑恶性脑膜瘤原代细胞,稳定传代后细胞免疫荧光鉴定;将细胞用立体定向仪接种于6只裸鼠大脑右侧顶叶,同时用注射器种植于6只裸鼠皮下;6周后处死裸鼠,HE染色和免疫组化检测肿瘤的增殖以及对周边脑组织的侵犯。结果成功培养并稳定传代恶性脑膜瘤原代细胞(术后病理:间变型恶性脑膜瘤),细胞免疫荧光检测Vimentin(+)和EMA(+),生长曲线呈"S"型,FITC法流式细胞术检测Q3区域占95.99±2.58%;6周后,皮下成瘤组可见明显肿瘤结节,原位成瘤组裸鼠明显消瘦;成瘤组织切片染色皮下成瘤组和颅内成瘤组Ki-67表达约36±5.35%,颅内成瘤组周边脑组织侵犯明显。结论成功培养人恶性脑膜瘤原代细胞并实现稳定传代,首次探究人恶性脑膜瘤原代细胞原位成瘤模型的建立。

Objective To obtain stable primary cultures of human malignant meningioma cells and establish an intracranial insitu tumor model in nude mice. Methods Ten surgical specimens of highly suspected malignant meningioma were obtained with postoperative pathological confirmation. Primary malignant meningioma cells were cultured from the tissues using a modified method and passaged. After identification with cell immunofluorescence, the cultured cells were inoculated into the right parietal lobe of 6 nude mice using stereotaxic apparatus and also transplanted subcutaneously in another 6 nude mice.The nude mice were executed after 6 weeks, and HE staining and immunohistochmistry were used to detect tumor growth and the invasion of the adjacent brain tissues. Results The primary malignant meningioma cells were cultured successfully,and postoperative pathology reported anaplastic malignant meningioma. Cell immunofluorescence revealed positivity for vimentin and EMA in the cells, which showed a S-shaped growth curve in culture. Flow cytometry revealed a cell percentage in the Q3 area of（95.99±2.58）%. Six weeks after transplantation, tumor nodules occurred in the subcutaneous tumor group, and the nude mice bearing the in situ tumor showed obvious body weight loss. The xenografts in both groups contained a mean of（36±5.35）% cells expressing Ki-67, and the intracranial in situ tumor showed obvious invasion of the adjacent peripheral brain tissues. Conclusion We obtained stable primary cultures of malignant meningioma cells and successfully established a nude mouse model bearing in situ human malignant meningioma.