空间环境对B16细胞生物学特性的影响

发布时间：2018-05-21 19:18

本文选题：空间环境 + B16细胞　；参考：《昆明医学院》2006年硕士论文

【摘要】：研究目的： 1．通过动物实验观察空间诱变小鼠黑色素瘤B16细胞株的成瘤性，探索通过体内实验筛选空间诱变B16细胞的方法，寻找免疫原性发生变异的阳性细胞株。 2．观察可疑阳性空间诱变小鼠黑色素瘤B16细胞细胞骨架和细胞表面结构，观察可疑变异细胞株的细胞骨架和表面结构与对照细胞株的差异。 3．检测体内筛选获得的实验阳性细胞株的体外培养生长曲线和细胞周期分布等生物学特性，观察可疑变异细胞株的生物学特性与对照细胞株的差异。 4．观察可疑变异细胞株基因表达的改变，寻找与黑色素瘤发生及转移密切相关的基因。研究方法： 1．将B16细胞株应用细胞低温长期生存系统，搭载于我国第20颗返回式卫星，返地后进行单克隆化，从得到的110株单克隆太空诱变B16细胞株中随机选取4株，常规培养传代5代后和对照细胞株同时分别接种C57BL/6J小鼠，腹腔接种组观察生存期，皮下接种组14天后取血、处死，记录瘤重，脾脏重，胸腺重，并进行血清细胞因子检测和瘤体的病理学检测。 2．应用激光共聚焦显微镜观察筛选所得阳性细胞株与对照细胞株分别用荧光素标记的鬼笔环肽染色，观察细胞骨架比较异同。 3．应用原子力显微镜(AFM)对筛选所得阳性细胞株和对照细胞株分别成像，分别观察空间诱变细胞株和对照组细胞所得图像的差异。 4．MTT法检测筛选所得阳性细胞株细胞生长曲线与对照组进行比较。 5．应用流式细胞仪(FCM)检测筛选所得阳性细胞株和对照细胞株的细胞周期分布。
[Abstract]:Objectives of the study: 1. The tumorigenesis of murine melanoma B16 cell line induced by space was observed through animal experiments. The method of screening space mutated B16 cell line in vivo was explored to find out the positive cell line with variant immunogenicity. 2. The cytoskeleton and surface structure of murine melanoma B16 cells induced by suspected positive space mutagenesis were observed, and the difference of cytoskeleton and surface structure between murine melanoma B16 cell line and control cell line was observed. 3. The biological characteristics of culture growth curve and cell cycle distribution in vitro were detected, and the differences between the cell line and the control cell line were observed. 4. To observe the changes of gene expression in suspected variant cell lines and to search for genes closely related to melanoma development and metastasis. Research methods: 1. The B16 cell line was carried on the 20th recoverable satellite in China with a cryogenic long-term survival system. After returning to earth, the B16 cell line was monoclonal transformed. Four of the 110 Monoclonal Space mutated B16 cell lines were randomly selected. C57BL/6J mice were inoculated with conventional culture for 5 passages and control cell lines at the same time. The survival time was observed in the intraperitoneal inoculation group. The blood was taken from the subcutaneous inoculation group after 14 days. The tumor weight, spleen weight, thymus weight were recorded, and the tumor weight, spleen weight and thymus weight were recorded. Serum cytokines and tumor pathology were detected. 2. Laser confocal microscopy was used to observe the difference of cytoskeleton between the positive cell line and the control cell line. 3. Atomic force microscopy (AFM) was used to image the positive cell lines and control cell lines, respectively, and to observe the difference between the space mutagenesis cell lines and the control cells. 4.MTT assay was used to detect the cell growth curve of positive cell line and to compare with the control group. 5. Flow cytometry (FCM) was used to detect the cell cycle distribution of positive cell lines and control cell lines.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R85

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