乳腺成体干细胞体外分离、培养、鉴定及药物干预研究

发布时间：2018-01-16 15:34

本文关键词：乳腺成体干细胞体外分离、培养、鉴定及药物干预研究　出处：《吉林大学》2006年硕士论文　论文类型：学位论文

【摘要】：乳腺干细胞应为成体干细胞的一种,其具有无限自我更新能力并能产生子代细胞的能力,本实验体外分离、培养并鉴定乳腺干细胞,观察其生长特征并进行药物干预研究。第一部分实验通过免疫磁株技术,对静止期乳腺的乳腺癌患者癌旁正常组织进行体外培养,并分离出乳腺上皮细胞:导管上皮细胞和肌上皮细胞。其中导管上皮细胞的一个亚群可以向肌上皮细胞转化,通过三维培养可以形成类似的乳腺终末导管结构。并通过免疫组化染色对分离的细胞亚群进行鉴定染色。通过脂质体转染的方法向乳腺上皮细胞内转入SV40质粒,并观察到可使乳腺上皮细胞体外多次传代而不死,获得永生化。并通过免疫磁株技术分离得到的导管上皮细胞亚群具有干细胞特性,而细胞形态无明显改变。第二部分实验对分离培养后的乳腺干细胞加入不同浓度的化疗药物氟尿嘧啶和紫杉醇,以及中药黄芪注射液和Rg3单体,观察其对乳腺干细胞生长的影响。本实验初步分离出具有成体干细胞特性的乳腺导管上皮细胞亚群,并对其细胞表面标记物进行免疫组化染色鉴定。对分离的乳腺干细胞加入药物,并筛选出对乳腺干细胞生长有抑制作用的化疗药物和对其生长有促进作用的中药。
[Abstract]:Breast stem cells should be a kind of adult stem cells. It has the ability of unlimited self renewal and can generate progeny cells. In vitro isolation, culture and identification of breast stem cells, observing the growth characteristics and drug intervention.
The first part of the experiment through the immune magnetic cell technology, breast cancer in resting stage breast adjacent normal tissues were cultured in vitro, and isolated mammary epithelial cells, ductal epithelial cells and myoepithelial cells. The ductal epithelial cells of a subgroup can be transformed to myoepithelial cells, through the three-dimensional culture can form similar breast terminal ductal structures. By immunohistochemical staining of isolated cell subsets were identified by staining. Into SV40 plasmid to mammary epithelial cells by liposome transfection method, and observed the mammary epithelial cells in vitro Subculture and die, immortalized. And through the immune magnetic cell separation technology by ductal epithelium subpopulations of cells with stem cell properties, but no obvious changes in cell morphology.
In the second part, the effects of different concentrations of chemotherapeutic drugs, fluorouracil and paclitaxel, and astragalus injection and Rg3 monomer on breast stem cells after culture were observed.
This study preliminary isolated characteristics of adult stem cells of breast ductal epithelial cell subsets, and the cell surface markers by immunohistochemical staining. The separation of breast stem cells into drugs, and screening of traditional Chinese medicine breast stem cells growth inhibitory effect of chemotherapeutic agents and to promote growth role.

【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329.2

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