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Study on the Effect of Mature Myocardial Cell to Induce the Early Embryo or ESCs Differentisted Directly

Author: JiangFangPing
Tutor: ZhangXiaoGang
School: Chongqing Medical University
Course: Internal Medicine
Keywords: Embryonic stem cells Myocardial cells Co-culture Cell differentiation
CLC: R329
Type: Master's thesis
Year: 2008
Downloads: 93
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Abstract

Prepared to substantiate the mature myocardial cells of early embryos or embryonic stem cells (ESCs) directed differentiation special induced to provide a theoretical basis for the establishment of a set of simple, practical embryonic stem cell transplantation technology. The method of separation of SD rats were cultured neonatal rat cardiomyocytes dye DAPI (4'6-associated squint -2 - phenylindole) nuclear cell labeling. The the applications naturally mated and the hyperstimulation technology to obtain the Kunming mice early embryos (8 to 16-cell stage embryos) co-cultured myocardial cells and myocardial microenvironment; take Kunming mice (3.5 to 4 days) after blastocyst culture isolated ESCs, 1-2 generations of cell mass and myocardial cells of mouse ESCs and cardiac fibroblasts co-cultured. Video dynamic observation of early embryos or ESCs differentiation into myocardial cells; respectively, 3, 7, and 14 days after co-culture line troponin T (cTnT), α-actin protein (α-Actinin) immunofluorescence staining. Results (1) early embryonic cardiomyocytes or cardiac fibroblasts co-culture, can continue development and differentiation, the differentiation of the cells cTnT expression. (2) a generation or two generations ESCs cells and / or cell mass and myocardial cells co-cultured rhythm beating cardiomyocyte-like cells in about seven days; observed up to 14 days, beating rate of approximately 30 to 50 times / min. When the The best experimental batches count ESCs cells more than 1/4 and / or ESCs cell mass rhythmic beats; ESCs cells and / or cell mass and myocardial cells co-cultured and add 0.6% DMSO, rhythm pulsating cardiomyocyte-like cells rate is not significantly improved. (3) myocardial cells induced group, record pulsatile and non-pulsatile ESCs cardiomyocyte-like cells induced differentiation of cTnT, α-actin protein staining positive rate was 27%; ESCs and cardiac fibroblasts co-cultured fibroblasts induced group, differentiated cells of cTnT, α-actin protein fluorescence staining were negative; about 3% of the differentiated cells cTnT 0.6% DMSO inducing group of α-actin protein fluorescence staining; DMSO combined myocardial cells induced group, the results with myocardial cells The induced group similar. Conclusions (1) cardiomyocytes and cardiac fibroblasts micro environment is conducive to early embryonic development and differentiation, differentiation of the cells can continue to develop proliferation. However, you can not induce the the cTnT positive expression of the myocardial-like cells, consider the major impact of the direct contact between the cell differentiation of trophoblast cells, leading to induction failure. (2) without the built system operation ESCs can be induced to differentiate for rhythm pulsating cardiomyocyte-like cells; mature cardiomyocytes ESCs co-cultured state, mature cardiomyocytes to myocardial cells can be induced ESCs ESCs to this microenvironment directed differentiation of myocardial cells was significantly higher than natural differentiation group that mature cardiomyocytes induced ESCs cells directed the strong differentiation induced factors.

Study on the Effect of Mature Myocardial Cell to Induce the Early Embryo or ESCs Differentisted Directly

Abstract

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