ZNF207基因在P19细胞诱导分化过程中表达水平的研究

发布时间：2018-05-15 11:26

本文选题：P19细胞 + 心肌细胞　；参考：《南京医科大学》2007年硕士论文

【摘要】： 【目的】体外培养P19细胞，并诱导P19细胞向胚胎心肌样细胞分化。【方法】P19细胞经复苏、培养、传代后，在体外二甲基亚砜(DMSO)诱导下向心肌细胞分化，在光镜、倒置显微镜下观察细胞形态学变化。【结果】复苏后未分化的P19细胞大小均匀、分散，胞体小，呈圆形，3～4 h开始有细胞贴壁，呈不规则扁平状，24h后细胞长满瓶底的50％左右，48h细胞生长密集。0.9％二甲基亚砜(DMSO)诱导4d，有细胞聚集体形成。诱导4d后，使细胞贴壁生长，以分散、克隆集落方式增殖。d8，可以看见心肌样搏动，随分化时间延长，细胞形态多样化，梭形细胞增多。【结论】P19细胞在0.9％DMSO诱导下已有向心肌细胞分化的趋势，有类似胚胎心肌样细胞形成。【目的】观察锌指蛋白207基因(ZNF207)在P19细胞向心肌细胞诱导分化过程中表达水平的变化，探讨ZNF207基因与心肌细胞分化之间可能的关系。【方法】体外培养P19细胞，诱导细胞向心肌细胞分化，采用RT-PCR技术在细胞分化成熟的不同时段检测P19细胞中ZNF207基因mRNA表达水平。【结果】ZNF207基因在P19细胞向心肌细胞分化的过程中均有表达，，随细胞分化成熟，该基因表达水平逐渐升高。经统计学分析发现，在分化第0～2天该基因表达无差异；第2～10天表达持续性增高，各时间点之间均有差异，差异均有统计学意义(P＜0.05)；第10～17天该基因稳定高表达，各时间点之间无差异。【结论】ZNF207基因在P19细胞向心肌细胞分化过程中表达逐渐上调，可能有利于心肌细胞的分化和发育。
[Abstract]:Objective: to culture P19 cells in vitro and induce P19 cells to differentiate into embryonic cardiomyocyte-like cells. [methods] after resuscitation, culture and passage, P19 cells differentiated into cardiomyocytes induced by dimethyl sulfoxide (DMSO) in vitro. Morphological changes of P19 cells were observed under light microscope and inverted microscope. [results] after resuscitation, the undifferentiated P19 cells were uniform in size, dispersed in size and small in size. After 24 hours of irregular flattening, 50% of the cells covered the bottom of the bottle. The cells grew densely. 0.9% DMSO (dimethyl sulfoxide) was induced for 4 days. After 4 days of induction, the cells were adherent to the wall and proliferated in a way of dispersing and clone colony. Myocardial beating could be seen. With the extension of the differentiation time, the morphology of the cells was diversified, and the spindle cells were increased. [conclusion] P19 cells have a tendency to differentiate into cardiomyocytes induced by 0.9%DMSO, which is similar to embryonic cardiomyocyte formation. [objective] to observe the expression of zinc finger protein 207 (ZNF207) in P19 cells and to explore the possible relationship between ZNF207 gene and cardiomyocyte differentiation. [methods] P19 cells were cultured in vitro and induced to differentiate into cardiomyocytes. RT-PCR technique was used to detect the expression of ZNF207 gene mRNA in P19 cells at different stages of cell differentiation and maturation. [results] ZNF207 gene was expressed during the differentiation of P19 cells into cardiomyocytes, and the expression level of ZNF207 gene increased gradually with the differentiation and maturation of P19 cells. Statistical analysis showed that there was no difference in the expression of the gene on the 0th day of differentiation, the expression of the gene continued to increase on the 10th day, and there were significant differences among all the time points (P < 0.05), and the expression of the gene was stable and high on the 10th day, 17 days after differentiation. There is no difference between time points. [conclusion] the expression of ZNF207 gene is up-regulated during the differentiation of P19 cells into cardiomyocytes, which may be beneficial to the differentiation and development of cardiomyocytes.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329

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