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目的:通过中断BMSCs的正常分化过程,探索肿瘤发生与干细胞分化受阻的关系。方法:全骨髓贴壁培养法获取BMSCs,β-巯基乙醇法诱导BMSCs分化为神经细胞,同时,在加入诱导培养基后的不同时间中断诱导,正常培养至第四代及之后。通过免疫组化检测、软琼脂克隆形成实验和CTX免疫抑制小鼠成瘤实验等鉴定和分析中断神经诱导细胞的分化特性和恶性转变趋势。结果:通过免疫组化检测中断神经诱导细胞逐渐表达了神经丝蛋白(NF);通过软琼脂集成形成实验,中断神经诱导40h组的细胞表现了比其他组较强的克隆形成能力,并在细胞病理学HE染色实验和细胞CTX免疫抑制小鼠成瘤实验证明中断神经诱导40h组的细胞发生了病理性恶变和获得致瘤性。结论:中断BMSCs的正常分化的培养,细胞有转化为肿瘤细胞的趋势。
OBJECTIVE: To explore the relationship between tumorigenesis and blocked stem cell differentiation by disrupting the normal differentiation process of BMSCs. Methods: BMSCs were obtained by whole bone marrow adherent culture method. BMSCs were induced to differentiate into neurons by β-mercaptoethanol method. At the same time, BMSCs were induced to differentiate at different time after inducing culture medium and were cultured to the fourth generation and later. Immunohistochemical detection, soft agar colony formation assay and CTX immunosuppressive mouse tumorigenicity experiment were used to identify and analyze the differentiation characteristics and malignant transformation tendency of nerve-induced cells. Results: Neurofilament protein (NF) was gradually expressed in the induced neurons by immunohistochemistry. Through the soft agar integration experiment, the cells induced by the interruption of neural induction in 40h showed stronger clonogenicity than the other groups, Histopathological HE staining and CTX immunosuppression mice tumorigenicity experiments showed that the nerve cells interrupted 40h induced pathological malignant transformation and tumorigenicity. Conclusion: The normal differentiation of BMSCs is interrupted and the cells have the tendency of transforming into tumor cells.