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人恶性脑膜瘤细胞的原代培养及其在裸鼠颅内原位移植

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

作者信息

Hu Mei-Xin, Liu Jia-le, Chen Xuan-Bo, Xu An-Qi, Shu Song-Ren, Wang Chao-Hu, Liu Yi

机构信息

First Clinical College, Southern Medical University, Guangzhou 510515, China. E-mail:

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2018 Mar 20;38(3):340-345. doi: 10.3969/j.issn.1673-4254.2018.03.16.

Abstract

OBJECTIVE

To obtain stable primary cultures of human malignant meningioma cells and establish an intracranial in-situ 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.

摘要

目的

获取人恶性脑膜瘤细胞的稳定原代培养物,并在裸鼠中建立颅内原位肿瘤模型。

方法

获取10例高度怀疑为恶性脑膜瘤的手术标本,术后经病理证实。采用改良方法从组织中培养原代恶性脑膜瘤细胞并传代。经细胞免疫荧光鉴定后,使用立体定向仪将培养的细胞接种到6只裸鼠的右顶叶,另6只裸鼠皮下移植。6周后处死裸鼠,采用苏木精-伊红(HE)染色和免疫组织化学检测肿瘤生长及对相邻脑组织的侵袭情况。

结果

成功培养出原代恶性脑膜瘤细胞,术后病理报告为间变性恶性脑膜瘤。细胞免疫荧光显示细胞中波形蛋白和上皮膜抗原(EMA)呈阳性,细胞在培养中呈S形生长曲线。流式细胞术显示Q3区域细胞百分比为(95.99±2.58)%。移植6周后,皮下肿瘤组出现肿瘤结节,原位肿瘤裸鼠体重明显减轻。两组异种移植瘤中平均有(36±5.35)%的细胞表达Ki-67,颅内原位肿瘤显示对相邻外周脑组织有明显侵袭。

结论

我们获得了恶性脑膜瘤细胞的稳定原代培养物,并成功建立了人恶性脑膜瘤原位裸鼠模型。

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