催产素诱导P19细胞分化为心肌细胞的实验研究

发布时间：2018-04-14 05:17

本文选题：催产素 + P19细胞　；参考：《河北医科大学》2006年硕士论文

【摘要】： 目的: 心肌梗死患者的干细胞移植治疗能够取代坏死的心肌细胞、增加有功能的心肌细胞数量,从而达到改善心功能的目的,是一种理想的治疗手段。目前,干细胞治疗还处于基础实验研究和初期的临床研究阶段,是全球心血管疾病研究者关注的焦点。P19胚胎癌细胞是目前研究较成熟的一种胚胎源性干细胞,深受广大干细胞研究者青睐。催产素(Oxytocin, OT)长期以来被认为只是一种女性生殖激素,然而,近年来随着其多方面作用的发现,逐渐被越来越多的研究人员所重视。近年的研究发现,OT还具有促进心脏发生的作用[1]。本实验应用OT作用于培养的P19胚胎源性干细胞,诱导其向心肌细胞方向分化,并通过免疫细胞化学技术、激光扫描共聚焦显微镜技术、透射电子显微镜技术等方法对诱导分化的细胞进行形态学观察,以期为心肌损伤干细胞治疗提供基础实验数据。方法: 1 P19细胞诱导分化取对数生长期的P19细胞,经0.25%胰酶-0.2%EDTA消化,制成单细胞悬液。调整细胞密度为2×10~5/ml,以每皿2ml接种于0.5%琼脂铺底的直径35mm细菌级培养皿中悬浮培养。实验组1、2、3的培养基中分别含OT 1×10~(-7) mol/ L、3×10~(-7) mol/ L、5×10~(-7) mol/ L;对照组的培养基中不含OT。
[Abstract]:Objective:Stem cell transplantation in patients with myocardial infarction can replace the necrotic cardiomyocytes and increase the number of functional cardiomyocytes so as to improve the cardiac function and is an ideal treatment method.At present, stem cell therapy is still in the stage of basic experimental research and early clinical research, which is the focus of global cardiovascular disease researchers. P19 embryonic cancer cell is a mature embryonic stem cell.It is favored by stem cell researchers.Oxytocin (OT) has long been considered as a female reproductive hormone. However, with the discovery of its various roles, more and more researchers pay more and more attention to it.In recent years, it has been found that OT can also promote the development of the heart [1].In this study, OT was used to induce the differentiation of P19 embryonic stem cells into cardiomyocytes, and by immunocytochemistry, laser scanning confocal microscopy (LSCM) was used to induce the differentiation of P19 embryonic stem cells into cardiomyocytes.Transmission electron microscopy (TEM) was used to observe the morphology of differentiated cells in order to provide experimental data for the treatment of myocardial injury stem cells.Methods:1 P19 cell differentiationP19 cells in logarithmic growth stage were digested by 0.25% trypsin-0.2TA and single cell suspension was prepared.The cell density was adjusted to 2 脳 10 ~ (5) / ml, and the suspension culture was carried out in 0.5% Agar (0.5% Agar) -coated diameter 35mm bacterial-grade culture dish with each dish of 2ml.OT1 脳 10 ~ (10) mol/ L ~ (7) mol/ L ~ (3 脳 10 ~ (-7)) mol/ L ~ (5 脳 10 ~ (-7)) mol/ L were found in the culture medium of the experimental group (1 脳 10 ~ (2) C), but not in the medium of the control group.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329

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