妊娠对脂肪干细胞增殖活性的影响
发布时间：2018-09-02 09:13
【摘要】：[目的]本实验采用组织工程学原理,提取不同妊娠时期的ADSCs进行培养、鉴定,然后对其增殖活性进行对比分析,同时检测雌孕激素对ADSCs增殖活性的影响。如实验结果符合预期,提取妊娠期干细胞作为种子细胞进行培养将可能取得良好效果,同时也是对其生物学特性的有益探索。 [方法](1)完成对ADSCs的分离培养,并适时对细胞进行传代,采用光镜观察细胞形态和状态；(2)通过对ADSCs表面抗原的检测及成骨诱导实验完成对ADSCs的鉴定；(3)选取不同分组的第3代ADSCs,通过MTT法和碘化丙锭(PI)染色分别检测各组吸光度值和细胞周期,绘制细胞生长曲线,并对检测结果进行统计学分析,对比不同组ADSCs增殖活性的差异。 [结果](1)各组细胞生长良好,呈梭形旋涡状增殖,组织分离24h后妊娠组贴壁的梭形细胞数量较未孕组多；(2)细胞表面抗原检测结果显示所培养细胞CD90+CD44+CD34-,成骨诱导结果显示有红色钙结节生成；(3)MTT实验结果显示：未孕ADSCs+P组与未孕ADSCs组,孕中期ADSCs组、临产ADSCs组、未孕ADSCs+E2组组间差异无统计学意义(P0.05)；孕中期ADSCs组、临产ADSCs组、未孕ADSCs+E2组与未孕ADSCs组相比差异有统计学意义(P0.05)；(4)PI染色检测各组细胞周期显示：孕中期ADSCs组,临产ADSCs组和普通ADSCs+E2组与普通ADSCs组相比,处于非增殖周期的Go/G1期细胞所占比例呈下降趋势,差异有统计学意义(p0.05)；未孕ADSCs+P组与未孕ADSCs组,孕中期ADSCs组与临产ADSCs组相比处于Go/G,期的细胞所占比例无明显增减,差异无统计学意义(p0.05)。 [结论]所培养的细胞为ADSCs,且在体外培养环境下生长良好；妊娠大鼠的ADSCs增殖活性较未妊娠者高,妊娠中期与临产期ADSCs增殖活性无明显差异；雌二醇能促进ADSCs的增殖,孕酮则未表现出促进ADSCs增殖的能力。
[Abstract]:[objective] to investigate the effects of estrogen and progesterone on the proliferation of ADSCs in different pregnancy stages by using tissue engineering principle, and then compared and analyzed its proliferative activity. If the experimental results are in line with the expectation, it is possible to obtain good results by extracting pregnancy stem cells as seed cells, and it is also a useful exploration of their biological characteristics. [methods] (1) the ADSCs was isolated and cultured, and the cells were subcultured at the right time. The morphology and state of the cells were observed by light microscope. (2) the ADSCs was identified by the detection of ADSCs surface antigen and the osteogenesis induction experiment. (3) the third generation ADSCs, of different groups were selected to detect the absorbance and cell cycle of each group by MTT method and (PI) staining respectively, and to draw the cell growth curve. The results were analyzed statistically to compare the proliferative activity of different groups of ADSCs. [results] (1) the cells in each group grew well and proliferated in fusiform swirl shape. After 24 hours of tissue separation, the number of fusiform cells adhered to the wall in pregnancy group was more than that in non-pregnant group. (2) the results of cell surface antigen test showed that the CD90 CD44 CD34-, osteogenesis of cultured cells showed red calcium nodule formation, (3) MTT test showed that: ADSCs P group, ADSCs group. There was no significant difference between non-pregnant ADSCs E2 group (P0.05), second trimester ADSCs group, parturient ADSCs group, non-pregnant ADSCs E2 group and non-pregnant ADSCs group had significant difference (P0.05); (4) PI staining showed that the second trimester ADSCs group, the non-pregnant ADSCs E2 group and the non-pregnant ADSCs group had significant difference (P0.05); (4). The percentage of Go/G1 cells in non-proliferative cycle in ADSCs group and ADSCs E2 group decreased significantly (p0.05), and ADSCs P group and non-pregnant ADSCs group showed significant difference (p0.05), the percentage of Go/G1 phase cells in non-proliferative cycle group was significantly lower than that in normal ADSCs group (p0.05), while that in non-pregnant ADSCs P group and non-pregnant ADSCs group was significantly lower (p0.05). The percentage of cells in Go/G, phase in ADSCs group was not significantly increased compared with that in parturient ADSCs group (p0.05). [conclusion] the cultured cells were ADSCs, and grew well in vitro, the proliferative activity of ADSCs in pregnant rats was higher than that in unpregnant rats, and the proliferative activity of ADSCs in the second trimester of pregnancy was not significantly different from that in parturient stage, estradiol could promote the proliferation of ADSCs. Progesterone did not show the ability to promote the proliferation of ADSCs.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329
本文编号：2218910
[Abstract]:[objective] to investigate the effects of estrogen and progesterone on the proliferation of ADSCs in different pregnancy stages by using tissue engineering principle, and then compared and analyzed its proliferative activity. If the experimental results are in line with the expectation, it is possible to obtain good results by extracting pregnancy stem cells as seed cells, and it is also a useful exploration of their biological characteristics. [methods] (1) the ADSCs was isolated and cultured, and the cells were subcultured at the right time. The morphology and state of the cells were observed by light microscope. (2) the ADSCs was identified by the detection of ADSCs surface antigen and the osteogenesis induction experiment. (3) the third generation ADSCs, of different groups were selected to detect the absorbance and cell cycle of each group by MTT method and (PI) staining respectively, and to draw the cell growth curve. The results were analyzed statistically to compare the proliferative activity of different groups of ADSCs. [results] (1) the cells in each group grew well and proliferated in fusiform swirl shape. After 24 hours of tissue separation, the number of fusiform cells adhered to the wall in pregnancy group was more than that in non-pregnant group. (2) the results of cell surface antigen test showed that the CD90 CD44 CD34-, osteogenesis of cultured cells showed red calcium nodule formation, (3) MTT test showed that: ADSCs P group, ADSCs group. There was no significant difference between non-pregnant ADSCs E2 group (P0.05), second trimester ADSCs group, parturient ADSCs group, non-pregnant ADSCs E2 group and non-pregnant ADSCs group had significant difference (P0.05); (4) PI staining showed that the second trimester ADSCs group, the non-pregnant ADSCs E2 group and the non-pregnant ADSCs group had significant difference (P0.05); (4). The percentage of Go/G1 cells in non-proliferative cycle in ADSCs group and ADSCs E2 group decreased significantly (p0.05), and ADSCs P group and non-pregnant ADSCs group showed significant difference (p0.05), the percentage of Go/G1 phase cells in non-proliferative cycle group was significantly lower than that in normal ADSCs group (p0.05), while that in non-pregnant ADSCs P group and non-pregnant ADSCs group was significantly lower (p0.05). The percentage of cells in Go/G, phase in ADSCs group was not significantly increased compared with that in parturient ADSCs group (p0.05). [conclusion] the cultured cells were ADSCs, and grew well in vitro, the proliferative activity of ADSCs in pregnant rats was higher than that in unpregnant rats, and the proliferative activity of ADSCs in the second trimester of pregnancy was not significantly different from that in parturient stage, estradiol could promote the proliferation of ADSCs. Progesterone did not show the ability to promote the proliferation of ADSCs.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329
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