【摘要】：目的： 探索小鼠MSC具有多分化潛能的原因以及MSCs對(duì)卵巢GC的影響。 方法： 1、從小鼠骨髓中分離MSCs,并進(jìn)行原代培養(yǎng),分別于0、2、4、6、8、10天提取總RNA,RT-PCR檢測(cè)多潛能標(biāo)記基因和相關(guān)因子、三個(gè)胚層標(biāo)記基因的表達(dá)情況。 2、利用分離、純化的MSCs與GC,建立共培養(yǎng)體系。實(shí)驗(yàn)分3組：?jiǎn)为?dú)培養(yǎng)MSCs組、單獨(dú)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(cè)卵巢內(nèi)標(biāo)記基因在GC和MSCs中的表達(dá)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(gè)胚層基因的表達(dá)情況。并在3天和5天通過MTT法檢測(cè)各組細(xì)胞的增殖情況。 結(jié)果： 小鼠MSCs表達(dá)多潛能性標(biāo)記基因Oct-4、nanog和相關(guān)因子Klf4和c-Myc.三個(gè)胚層基因nestin、SM22α、CYP51均有表達(dá)。在共培養(yǎng)實(shí)驗(yàn)中,單獨(dú)培養(yǎng)和共培養(yǎng)GC均有ZP1.ZP2.ZP3和FSHR的表達(dá),但ZP3.FSHR和BMP-15表達(dá)量不同。MSCs可促進(jìn)GC增殖。不同天數(shù)單獨(dú)培養(yǎng)和共培養(yǎng)MSCs均未檢測(cè)到卵巢內(nèi)標(biāo)記基因的表達(dá),但多能性基因和相關(guān)因子表達(dá)量不同,三個(gè)胚層基因表達(dá)無變化。此外,GC可促進(jìn)MSCs增殖。 結(jié)論： 貼壁篩選得到的小鼠MSCs表達(dá)多能性標(biāo)記基因以及三個(gè)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(jìn)增殖并影響基因的表達(dá)。
[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(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R329

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