發(fā)布時(shí)間：2018-05-15 11:26

  本文選題：P19細(xì)胞 + 心肌細(xì)胞�。� 參考：《南京醫(yī)科大學(xué)》2007年碩士論文

【摘要】： 【目的】體外培養(yǎng)P19細(xì)胞，并誘導(dǎo)P19細(xì)胞向胚胎心肌樣細(xì)胞分化。 【方法】P19細(xì)胞經(jīng)復(fù)蘇、培養(yǎng)、傳代后，在體外二甲基亞砜(DMSO)誘導(dǎo)下向心肌細(xì)胞分化，在光鏡、倒置顯微鏡下觀察細(xì)胞形態(tài)學(xué)變化。 【結(jié)果】復(fù)蘇后未分化的P19細(xì)胞大小均勻、分散，胞體小，呈圓形，3～4 h開始有細(xì)胞貼壁，呈不規(guī)則扁平狀，24h后細(xì)胞長滿瓶底的50％左右，48h細(xì)胞生長密集。0.9％二甲基亞砜(DMSO)誘導(dǎo)4d，有細(xì)胞聚集體形成。誘導(dǎo)4d后，使細(xì)胞貼壁生長，以分散、克隆集落方式增殖。d8，可以看見心肌樣搏動(dòng)，隨分化時(shí)間延長，細(xì)胞形態(tài)多樣化，梭形細(xì)胞增多。 【結(jié)論】P19細(xì)胞在0.9％DMSO誘導(dǎo)下已有向心肌細(xì)胞分化的趨勢，有類似胚胎心肌樣細(xì)胞形成。 【目的】觀察鋅指蛋白207基因(ZNF207)在P19細(xì)胞向心肌細(xì)胞誘導(dǎo)分化過程中表達(dá)水平的變化，探討ZNF207基因與心肌細(xì)胞分化之間可能的關(guān)系。 【方法】體外培養(yǎng)P19細(xì)胞，誘導(dǎo)細(xì)胞向心肌細(xì)胞分化，采用RT-PCR技術(shù)在細(xì)胞分化成熟的不同時(shí)段檢測P19細(xì)胞中ZNF207基因mRNA表達(dá)水平。 【結(jié)果】ZNF207基因在P19細(xì)胞向心肌細(xì)胞分化的過程中均有表達(dá)，，隨細(xì)胞分化成熟，該基因表達(dá)水平逐漸升高。經(jīng)統(tǒng)計(jì)學(xué)分析發(fā)現(xiàn)，在分化第0～2天該基因表達(dá)無差異；第2～10天表達(dá)持續(xù)性增高，各時(shí)間點(diǎn)之間均有差異，差異均有統(tǒng)計(jì)學(xué)意義(P＜0.05)；第10～17天該基因穩(wěn)定高表達(dá)，各時(shí)間點(diǎn)之間無差異。 【結(jié)論】ZNF207基因在P19細(xì)胞向心肌細(xì)胞分化過程中表達(dá)逐漸上調(diào)，可能有利于心肌細(xì)胞的分化和發(fā)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.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 黃仲賢，顧偉強(qiáng)，胡紅雨;鋅指類基因調(diào)控蛋白──生物無機(jī)化學(xué)和分子生物學(xué)發(fā)展的新領(lǐng)域[J];生物化學(xué)與生物物理進(jìn)展;1995年03期

2 錢玲梅,曹克將,黃元鑄,孔祥清,單其俊;房間隔缺損患者心肌組織基因表達(dá)譜系特征的初步分析[J];中華心血管病雜志;2003年03期

本文編號(hào)：1892259