本文關(guān)鍵詞：雌性大鼠骨髓干細(xì)胞離體從頭合成雌二醇的研究　出處：《南昌大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 骨髓干細(xì)胞 分化 性激素生成細(xì)胞 芳香化酶 雌二醇

【摘要】： 目的: 研究雌性大鼠骨髓干細(xì)胞(female bone marrow stem cells, FBMSCs)在含10%胎牛血清的高糖DMEM離體培養(yǎng)條件下細(xì)胞形態(tài)的變化,檢測(cè)能否分化為產(chǎn)生雌二醇(estradiol,E2)及表達(dá)芳香化酶(aromatase, P450arom)的性激素生成細(xì)胞,為卵巢早衰、老年人骨質(zhì)疏松癥等疾病的治療提供新的可能途徑。 方法: 全骨髓貼壁法分離成年雌性SD大鼠的骨髓干細(xì)胞,用含10%胎牛血清的高糖DMEM培養(yǎng)增殖,傳代至第3代,加或不加全反式視黃酸(all-trans retinoic acid, ATRA)誘導(dǎo)分化,分為對(duì)照組(不加ATRA)和ATRA組(加ATRA 10-5 mol/L),觀察兩組細(xì)胞的形態(tài)學(xué)變化;放射免疫法(radioimmunoassay, RIA)/酶聯(lián)免疫吸附法(enzyme-linked immunosorbent assay, ELISA)檢測(cè)兩組細(xì)胞培養(yǎng)液中E2、睪酮(testosterone, T)/雄烯二酮(androstenedione,ASD)的濃度;RT-PCR檢測(cè)兩組細(xì)胞中芳香化酶基因CYP19 mRNA的表達(dá)和FSHR mRNA的表達(dá);免疫細(xì)胞化學(xué)檢測(cè)兩組細(xì)胞中芳香化酶蛋白的表達(dá)情況。 結(jié)果: 1.離體培養(yǎng)的第3代雌性大鼠骨髓干細(xì)胞,主要表現(xiàn)為大小均一、梭形的貼壁細(xì)胞。 2. DMEM高糖培養(yǎng)基加胎牛血清培養(yǎng)第3代雌性大鼠骨髓干細(xì)胞,RT-PCR示對(duì)照組和ATRA組從培養(yǎng)第1天開(kāi)始均表達(dá)合成雌二醇的關(guān)鍵酶芳香化酶CYP19 mRNA和顆粒細(xì)胞標(biāo)志基因FSHR mRNA,免疫細(xì)胞化學(xué)示兩組細(xì)胞從培養(yǎng)第2天到第4天都有芳香化酶蛋白的表達(dá),陽(yáng)性細(xì)胞呈簇狀分布,形態(tài)呈多角形或梭形,細(xì)胞核呈卵圓形,染深藍(lán)色。從培養(yǎng)1天開(kāi)始在兩組細(xì)胞培養(yǎng)液中都檢測(cè)到較高水平的E2。 3.對(duì)照組和ATRA組都能產(chǎn)生T和ASD,提示雌性大鼠骨髓干細(xì)胞在離體培養(yǎng)條件下能從頭合成E2。 結(jié)論: 雌性大鼠骨髓干細(xì)胞在離體培養(yǎng)條件下能從頭合成E2及表達(dá)芳香化酶,分化為性激素生成細(xì)胞。
[Abstract]:Objective: To study female bone marrow stem cells of female rat bone marrow stem cells. The morphological changes of high glucose DMEM containing 10% fetal bovine serum were observed in vitro to determine whether the cells could differentiate into estradiol producing estradiol. E2) and the expression of aromatase, The sex hormone producing cells of P450 aromb provide a new way for the treatment of premature ovarian failure and osteoporosis in the elderly. Methods: Bone marrow stem cells from adult female SD rats were isolated by whole bone marrow adherent method. The stem cells were cultured and proliferated with high glucose DMEM containing 10% fetal bovine serum. Differentiation was induced by adding or without all-trans retinoic acid (ATRA). The cells were divided into two groups: the control group (without ATRAA) and the ATRA group (with ATRA 10-5 mol / L). The morphological changes of the two groups were observed. Radioimmunoassay. Enzyme linked immunosorbent assay (Elisa) was used to detect E _ 2 in the culture medium of the two groups. Testosterone testosterone, TX / androstenedione ASD; The expression of aromatase gene CYP19 mRNA and FSHR mRNA were detected by RT-PCR. The expression of aromatase protein was detected by immunocytochemistry. Results: 1. The third generation female rat bone marrow stem cells cultured in vitro mainly showed uniform size and fusiform adherent cells. 2. The third generation female rat bone marrow stem cells were cultured in DMEM high sugar medium and fetal bovine serum. RT-PCR showed that both the control group and the ATRA group expressed CYP19 mRNA, the key enzyme of estradiol synthesis, and the granulosa cell marker gene FSHR mRNA from the first day of culture. Immunocytochemistry showed that aromatase protein was expressed in the two groups from the second day to the fourth day of culture. The positive cells were distributed in clusters with the shape of polygonal or fusiform and the nucleus was oval. High levels of E2 were detected in both groups of cell cultures from the first day of culture. 3. Both control group and ATRA group can produce T and ASD, suggesting that female rat bone marrow stem cells can synthesize E2 from scratch in vitro. Conclusion: Female rat bone marrow stem cells can synthesize E2 and express aromatase from scratch in vitro and differentiate into sex hormone producing cells.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R329

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 倪秀麗;雌鼠骨髓干細(xì)胞懸滴培養(yǎng)形成卵泡樣結(jié)構(gòu)及性激素合成相關(guān)基因表達(dá)探討[D];南昌大學(xué);2011年

，

本文編號(hào)：1384805