發(fā)布時間：2018-12-26 11:33

【摘要】：目的： 探索小鼠MSC具有多分化潛能的原因以及MSCs對卵巢GC的影響。 方法： 1、從小鼠骨髓中分離MSCs,并進行原代培養(yǎng),分別于0、2、4、6、8、10天提取總RNA,RT-PCR檢測多潛能標(biāo)記基因和相關(guān)因子、三個胚層標(biāo)記基因的表達情況。 2、利用分離、純化的MSCs與GC,建立共培養(yǎng)體系。實驗分3組：單獨培養(yǎng)MSCs組、單獨培養(yǎng)GC組及MSCs和GC共培養(yǎng)組。分別在4天和6天提取總RNA,RT-PCR檢測卵巢內(nèi)標(biāo)記基因在GC和MSCs中的表達情況,以及共培養(yǎng)MSCs多潛能標(biāo)記基因和相關(guān)因子及三個胚層基因的表達情況。并在3天和5天通過MTT法檢測各組細胞的增殖情況。 結(jié)果： 小鼠MSCs表達多潛能性標(biāo)記基因Oct-4、nanog和相關(guān)因子Klf4和c-Myc.三個胚層基因nestin、SM22α、CYP51均有表達。在共培養(yǎng)實驗中,單獨培養(yǎng)和共培養(yǎng)GC均有ZP1.ZP2.ZP3和FSHR的表達,但ZP3.FSHR和BMP-15表達量不同。MSCs可促進GC增殖。不同天數(shù)單獨培養(yǎng)和共培養(yǎng)MSCs均未檢測到卵巢內(nèi)標(biāo)記基因的表達,但多能性基因和相關(guān)因子表達量不同,三個胚層基因表達無變化。此外,GC可促進MSCs增殖。 結(jié)論： 貼壁篩選得到的小鼠MSCs表達多能性標(biāo)記基因以及三個胚層標(biāo)記基因。在共培養(yǎng)體系中,GC與MSCs之間可以相互促進增殖并影響基因的表達。
[Abstract]:Objective: to explore the reasons for the polydifferentiation potential of mouse MSC and the effect of MSCs on ovarian GC. Methods: 1. MSCs, was isolated from the bone marrow of mice and cultured in primary culture. The total RNA,RT-PCR was extracted for detection of multipotential marker genes and related factors and the expression of the three genes in the embryo layer. 2. Co-culture system was established by separating and purifying MSCs and GC,. The experiment was divided into three groups: MSCs group, GC group and MSCs and GC co-culture group. Total RNA,RT-PCR was extracted on day 4 and day 6 to detect the expression of marker genes in GC and MSCs, and the expression of multipotential marker genes, related factors and three genes in the embryo layer of co-cultured MSCs. The proliferation of cells in each group was detected by MTT assay on 3 and 5 days. Results: mouse MSCs expressed multipotent marker gene Oct-4,nanog and related factors Klf4 and c-Myc. All three genes nestin,SM22 偽 and CYP51 were expressed. In the co-culture experiment, both ZP1.ZP2.ZP3 and FSHR were expressed in GC cultured alone and co-cultured, but the expression levels of ZP3.FSHR and BMP-15 were different. MSCs could promote the proliferation of GC. The expression of marker genes in ovary was not detected in MSCs cultured alone and cocultured for different days, but the expression levels of multipotent genes and related factors were different, and the expression of three genes in the embryo layer had no change. In addition, GC can promote the proliferation of MSCs. Conclusion: mouse MSCs expressed pluripotent marker gene and three blastocyst marker genes. In co-culture system, GC and MSCs can promote the proliferation of each other and affect gene expression.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R329

【參考文獻】

相關(guān)期刊論文 前9條

1 靳玉甫;覃愛平;;卵巢顆粒細胞P450芳香化酶研究進展[J];廣西醫(yī)科大學(xué)學(xué)報;2008年06期

2 魏素菊;張凱;;C-myc蛋白結(jié)構(gòu)、功能及對腫瘤細胞調(diào)控作用[J];河北醫(yī)藥;2009年21期

3 謝妍;乜春城;曹佐武;;小鼠ZP2 mRNA在卵泡顆粒細胞層的表達[J];暨南大學(xué)學(xué)報(自然科學(xué)與醫(yī)學(xué)版);2010年02期

4 賀衛(wèi)衛(wèi);謝琪璇;秦俊文;黃偉;李威娜;張春雪;肖鑾娟;;骨髓間充質(zhì)干細胞對損傷卵巢的修復(fù)作用[J];暨南大學(xué)學(xué)報(自然科學(xué)與醫(yī)學(xué)版);2010年02期

5 ;The signal transduction pathways and molecules for ES cells self-renewal[J];Chinese Science Bulletin;2005年08期

6 盧翠玲;楊巍;胡召元;劉以訓(xùn);;顆粒細胞的增殖分化及其在卵泡發(fā)育中的作用[J];科學(xué)通報;2005年21期

7 張鵬;石玉華;陳子江;;生長分化因子-9及生長分化因子9B/骨形態(tài)發(fā)生蛋白-15與卵巢功能[J];生殖與避孕;2007年03期

8 ;Identification of two distinct transactivation domains in the pluripotency sustaining factor nanog[J];Cell Research;2003年06期

9 鄧景岳;畢明剛;;影響原癌基因c-myc作用因素的研究進展[J];腫瘤;2009年12期

，

本文編號：2392067