The Research Of Transplanting The Human Embryonic Germ Cells To Treat The Acute Myocardial Infarction Of The Pig

Objectives:1.To establish the model of acute myocardial infarction in the pig;2.To observe the differentiation and repair function of human embryonic germ cells in the infarcted pig heart;3.To investigate the expression and significance of VEGF after transplanting human EG cell into the myocardium.Methods: Through tissue culture, the human EG cells were obtained by taking the gonadal ridges from human embryos aged 5～10 weeks. EG cells were isolated and cultured, at the same time, the human EG cells and human embryonic myocardial cells were cocultured to simulate cardiac micro-environment in vivo. The expression level of VEGF in culture supernatants on the third day and the ninth day of the both groups was detected respectively. The acute myocardial infarction (AMI) model was established by ligating the left anterior descending coronary artery in two pigs and they were divided into two groups:the transplanted group(hEGCs were injected into myocardium) and the control group(only PBS was injected). These pigs were killed respectively at the end of the second week after transplantation. Immunohistochemistry was used to detect the expression of anti–human cellular nuclei MAB1281 in order to locate the implanted cells and the expressions of transcription factor GATA-4 and connexin Cx43 were examined in the transplanted cells simultaneously.Results: A large number of human EG cells were acquired in vitro.The supernatant VEGF levels on the third day and the ninth day in the human EG cells and myocardial cells co-culture group were higher than the human EG cells group and the difference between the two groups was statistically significant (P<0.05); Typical ECG changes of myocardial infarction after coronary artery ligation showed that the acute myocardial infarction (AMI) model of pigs was successfully established ; In the transplanted group, transcription factor GATA-4 and connexin Cx43 showed positive presentation in the anti-human nuclei antibody MAB1281-positive cells, the control group was negative.Conclusion: Human EG cells transplanted into the myocardium of the pig through epicardium stay alive and differentiate into cardiomyocytes.The implanted Human EG cells can secrete cytokines such as VEGF to promote the repair of infarcted myocardium.