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目的研究体外培养的成人骨髓源性神经干细胞的致瘤性。方法对骨髓源性神经干细胞分别进行细胞形态学观察、刀豆球蛋白A凝集试验和双层软琼脂培养以探明其是否具有恶性转化细胞的形态特征、表面结构及生长特性的变化;利用免疫细胞化学的方法检测骨髓源性神经干细胞的端粒酶和肿瘤相关基因的表达;将骨髓源性神经干细胞接种到裸鼠体内观察其成瘤性。结果骨髓源性神经干细胞不具有恶性转化细胞的形态特征,在不同刀豆球蛋白A浓度下均未见明显的凝集反应,在双层软琼脂不能形成细胞克隆;骨髓源性神经干细胞的c-myc、c-fos和p53基因均呈阴性表达,而端粒酶逆转录酶呈弱阳性表达;将骨髓源性神经干细胞接种于裸鼠皮下6个月未见肿瘤形成,亦未见其它组织形成。结论骨髓源性神经干细胞保持了正常细胞的生物学特征,体内和体外的各项指标均未提示其具有致瘤性,体外的培养条件没有使其发生恶性转化,从致瘤性方面证实了骨髓源性神经干细胞临床移植的安全性。
Objective To study the tumorigenicity of adult human bone marrow-derived neural stem cells cultured in vitro. Methods Morphological observation, Concanavalin A agglutination test and double soft agar culture were used to investigate the morphological characteristics, surface structure and growth characteristics of bone marrow-derived neural stem cells. Immunohistochemistry Cytochemistry was used to detect the expression of telomerase and tumor-associated genes in bone marrow-derived neural stem cells. Bone marrow-derived neural stem cells were seeded into nude mice to observe their tumorigenicity. RESULTS: Bone marrow-derived neural stem cells did not have morphological features of malignant transformed cells. No obvious agglutination reaction was observed at different concanavalin A concentration, and cell clones could not be formed in bilayer soft agar. The c- myc, c-fos and p53 were all negatively expressed, whereas telomerase reverse transcriptase was weakly positive. There was no tumor formation and no other tissue formation was observed after subcutaneous inoculation of bone marrow-derived neural stem cells into nude mice for 6 months . Conclusion Bone marrow-derived neural stem cells maintain the biological characteristics of normal cells, and none of the indicators in vivo and in vitro suggest that they have tumorigenicity. In vitro culture conditions did not cause malignant transformation, and bone marrow Clinical neural stem cell transplantation safety.