乳腺成體干細(xì)胞體外分離、培養(yǎng)、鑒定及藥物干預(yù)研究
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本文關(guān)鍵詞:乳腺成體干細(xì)胞體外分離、培養(yǎng)、鑒定及藥物干預(yù)研究 出處:《吉林大學(xué)》2006年碩士論文 論文類型:學(xué)位論文
更多相關(guān)文章: 乳腺 成體干細(xì)胞 免疫磁化分離技術(shù) MUC ESA ASMA
【摘要】:乳腺干細(xì)胞應(yīng)為成體干細(xì)胞的一種,其具有無(wú)限自我更新能力并能產(chǎn)生子代細(xì)胞的能力,本實(shí)驗(yàn)體外分離、培養(yǎng)并鑒定乳腺干細(xì)胞,觀察其生長(zhǎng)特征并進(jìn)行藥物干預(yù)研究。 第一部分實(shí)驗(yàn)通過(guò)免疫磁株技術(shù),對(duì)靜止期乳腺的乳腺癌患者癌旁正常組織進(jìn)行體外培養(yǎng),并分離出乳腺上皮細(xì)胞:導(dǎo)管上皮細(xì)胞和肌上皮細(xì)胞。其中導(dǎo)管上皮細(xì)胞的一個(gè)亞群可以向肌上皮細(xì)胞轉(zhuǎn)化,通過(guò)三維培養(yǎng)可以形成類似的乳腺終末導(dǎo)管結(jié)構(gòu)。并通過(guò)免疫組化染色對(duì)分離的細(xì)胞亞群進(jìn)行鑒定染色。通過(guò)脂質(zhì)體轉(zhuǎn)染的方法向乳腺上皮細(xì)胞內(nèi)轉(zhuǎn)入SV40質(zhì)粒,并觀察到可使乳腺上皮細(xì)胞體外多次傳代而不死,獲得永生化。并通過(guò)免疫磁株技術(shù)分離得到的導(dǎo)管上皮細(xì)胞亞群具有干細(xì)胞特性,而細(xì)胞形態(tài)無(wú)明顯改變。 第二部分實(shí)驗(yàn)對(duì)分離培養(yǎng)后的乳腺干細(xì)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(duì)乳腺干細(xì)胞生長(zhǎng)的影響。 本實(shí)驗(yàn)初步分離出具有成體干細(xì)胞特性的乳腺導(dǎo)管上皮細(xì)胞亞群,并對(duì)其細(xì)胞表面標(biāo)記物進(jìn)行免疫組化染色鑒定。對(duì)分離的乳腺干細(xì)胞加入藥物,并篩選出對(duì)乳腺干細(xì)胞生長(zhǎng)有抑制作用的化療藥物和對(duì)其生長(zhǎng)有促進(jìn)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(xué)位授予單位】:吉林大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2006
【分類號(hào)】:R329.2
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