Progress In The Research Of Macaque Pluripotent Stem Cells And Fibroblasts Build Cardiomyocytes

Embryonic stem cells is an undifferentiated cell that separated from early embryos or primordial germ cells. iPSCs are adult cells that have been genetically reprogrammed to an embryonic stem cell-like state by being forced to express genes and factors important for maintaining the defining properties of embryonic stem cells. ESCs and iPSCs have a very important role in scientific progress and human health. Cardiovascular diseases are one of the leading causes of human death in the whole world. Symptomatic treatment can only relieve suffering for a while but never to cure the disease. Nonetheless, limited myocyte turnover does in fact occur in the adult mammalian heart; however, this is insufficient to restore contractile function following injury. We had to prompt methodological developments for creating de novo cardiomyocytes, both in vitro and in vivo. Currently mouse and rat were used in cardiomyocytes transplanted model, but mice and humans exhibit different sizes of myocytes, different myocardial capillary densities, and differing proportions of β-myosin versus a-myosin subunits within the ventricle. Functional effects of cellular cardiomyoplasty have to be found in animals that similar to human beings. So establishment non-human primates PSCs differentiate into myocardial cells and somatic cell transdifferented into cardiomyocytes has become an important key to understand this scientific issue.The experiment has two parts:the macaque somatic cell transdifferented into cardiomyocytes and PSCs differentiate into cardiomyocytes. The main contents of the first part is that constructing GATA4plasmid and introducing GATA4, OCT4, SOX2, KLF4plasmids into macaque cells, trying to turn macaque cells into cardiomyocytes. The main contents of the second part is that using two strains of monkeys PSCs including embryonic stem cell lines3-12and induced pluripotent stem cell lines iPS01.1, forming embryoid bodies in suspension culture and adding different concentrations of Vc or CH, geting the best differented system. Under the optimal differentiation system, comparing the beating cardiomyocytes ratio and the ratio of the beating numbers in the same day between3-12and iPS01.1. The results showed that (1) the differentiation of these groups all induced beating cardiomyocytes;(2) Beating cardiomyocytes occured at7days by adding20%serum differentiate and8days by adding10%serum indicated that serum promote differentiation of early cardiomyocytes;(3) ES3-12differentiated into cardiomyocytes ratio were:10+vc (1.4%);20+vc (7%);10+ch (0%);20+ch (30%) indicating that20%of serum with CH is the best differented system;(4) iPS01.1differentiated into cardiomyocytes ratio were:10+vc (1.6%);20+vc (4.3%);10+ch (1.25%);20+ch (41%) indicating that20%of serum with CH is the best differented system;(5) Under the best differented system iPSCs01.1differentiate into myocardial’s ratio is higher than the ES3-12;(6) Cardiomyocytes average beating frequency induced from iPS01.1in the4days is39/min, in the10days is21/min, Cardiomyocytes average beating frequency induced from ES3-12in the4days is63/min, in the10days is80/min indicating that cardiomyocytes induced from ES3-12is better than iPSCs01.1in beating frequency;(7) the differentiation of cardiomyocytes’s beat frequency become slowly with the time goes on.