小鼠MSCs多能性及其与颗粒细胞相互影响的研究
发布时间：2018-12-26 11:33
【摘要】：目的： 探索小鼠MSC具有多分化潜能的原因以及MSCs对卵巢GC的影响。 方法： 1、从小鼠骨髓中分离MSCs,并进行原代培养,分别于0、2、4、6、8、10天提取总RNA,RT-PCR检测多潜能标记基因和相关因子、三个胚层标记基因的表达情况。 2、利用分离、纯化的MSCs与GC,建立共培养体系。实验分3组：单独培养MSCs组、单独培养GC组及MSCs和GC共培养组。分别在4天和6天提取总RNA,RT-PCR检测卵巢内标记基因在GC和MSCs中的表达情况,以及共培养MSCs多潜能标记基因和相关因子及三个胚层基因的表达情况。并在3天和5天通过MTT法检测各组细胞的增殖情况。 结果： 小鼠MSCs表达多潜能性标记基因Oct-4、nanog和相关因子Klf4和c-Myc.三个胚层基因nestin、SM22α、CYP51均有表达。在共培养实验中,单独培养和共培养GC均有ZP1.ZP2.ZP3和FSHR的表达,但ZP3.FSHR和BMP-15表达量不同。MSCs可促进GC增殖。不同天数单独培养和共培养MSCs均未检测到卵巢内标记基因的表达,但多能性基因和相关因子表达量不同,三个胚层基因表达无变化。此外,GC可促进MSCs增殖。 结论： 贴壁筛选得到的小鼠MSCs表达多能性标记基因以及三个胚层标记基因。在共培养体系中,GC与MSCs之间可以相互促进增殖并影响基因的表达。
[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 伪 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.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329
本文编号：2392067
[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 伪 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.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329
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