本文關鍵詞：乳腺成體干細胞體外分離、培養(yǎng)、鑒定及藥物干預研究　出處：《吉林大學》2006年碩士論文　論文類型：學位論文

  更多相關文章： 乳腺 成體干細胞 免疫磁化分離技術 MUC ESA ASMA

【摘要】：乳腺干細胞應為成體干細胞的一種,其具有無限自我更新能力并能產生子代細胞的能力,本實驗體外分離、培養(yǎng)并鑒定乳腺干細胞,觀察其生長特征并進行藥物干預研究。 第一部分實驗通過免疫磁株技術,對靜止期乳腺的乳腺癌患者癌旁正常組織進行體外培養(yǎng),并分離出乳腺上皮細胞:導管上皮細胞和肌上皮細胞。其中導管上皮細胞的一個亞群可以向肌上皮細胞轉化,通過三維培養(yǎng)可以形成類似的乳腺終末導管結構。并通過免疫組化染色對分離的細胞亞群進行鑒定染色。通過脂質體轉染的方法向乳腺上皮細胞內轉入SV40質粒,并觀察到可使乳腺上皮細胞體外多次傳代而不死,獲得永生化。并通過免疫磁株技術分離得到的導管上皮細胞亞群具有干細胞特性,而細胞形態(tài)無明顯改變。 第二部分實驗對分離培養(yǎng)后的乳腺干細胞加入不同濃度的化療藥物氟尿嘧啶和紫杉醇,以及中藥黃芪注射液和Rg3單體,觀察其對乳腺干細胞生長的影響。 本實驗初步分離出具有成體干細胞特性的乳腺導管上皮細胞亞群,并對其細胞表面標記物進行免疫組化染色鑒定。對分離的乳腺干細胞加入藥物,并篩選出對乳腺干細胞生長有抑制作用的化療藥物和對其生長有促進作用的中藥。
[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.

【學位授予單位】：吉林大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R329.2

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本文編號：1433738