Role of Cardiac Transcription Factor Nkx2.5 on Cardiomyoplasty Model in vitro.
- Author:
Eun Kyung SHIN
1
;
Jeong Hyun PARK
;
Dae Joong KIM
;
Jang Hee HAHN
;
Kyeong Han PARK
;
Hoon Ki SUNG
;
Joo Young KIM
;
In Hwan SONG
;
Eon Gi SUNG
;
Yung Chang LEE
Author Information
1. Department of Anatomy, College of Medicine, Kangwon National University, Korea. jhpark@kangwon.ac.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
Cell transplantation;
P19;
Cardiomyocyte;
Nkx2.5;
Co-culture;
Intercellular junction
- MeSH:
Anoxia;
Cadherins;
Cardiomyopathies;
Cardiomyoplasty*;
Cell Survival;
Cell Transplantation;
Coculture Techniques;
Connexin 43;
Embryonal Carcinoma Stem Cells;
Intercellular Junctions;
Microscopy, Electron;
Myocardial Ischemia;
Myocytes, Cardiac;
Myofibrils;
Plasmids;
Prevalence;
Stem Cells;
Transcription Factors*;
Transfection;
Transplants
- From:Korean Journal of Anatomy
2004;37(1):31-41
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
Despite therapeutic advance, the prevalence of ischemic heart disease continues to increase. Recently, cell transplantation of stem cell has been proposed as a strategy for cardiac repair following myocardial damage. However, low differentiation efficiency into cardiomyocyte and poor cell viability associated with transplantation have limited the reparative capacity of these cell. In this study, we engineered P19 embryonal carcinoma cells using plasmid vector to overexpress the transcription factor MEF2c, Nkx2.5 involved in cardiomyogenesis. We investigated 1) formation of intercellular junction of P19 in mono-culture and co-culture with cardiomyocyte for functional and structural synchronous contraction after transplantation, 2) differentiation into cardiomyocyte, 3) resistance to hypoxic condition. An P19 embryonal carcinoma cell line expressing GFP, MEF2c, Nkx2.5 was generated by gene transfection and clonal selection. Nkx2.5 overexpression induced connexin43 expression level decrease. Electron microscopy revealed myofibril organization and immunostaining with cTnT showed positive staining in P19-Nkx2.5, consistent with early stage cardiomyocyte. Connexin43 and N-cadherin was expressed between P19-MEF2c and cardiomyocyte, P19- Nkx2.5 and cardiomyocyte in co-culture. And beating rate of cardiomyocyte co-cultured with P19-Nkx2.5 increased much more than other group, even if P19-Nkx2.5 did not have synchronous contraction with cardiomyocyte. Additionally, P19-Nkx2.5 had a resistance against hypoxia. These result suggest that overexpression of Nkx2.5 induced differentiation of P19 into cardiomyocyte and would be electro-mechanical coupling with cardiomyocyte after transplantation. Futhermore, Nkx2.5 overexpression had protection potential to hypoxic injury. Therefore, P19 cell overexpressed Nkx2.5 would be promising cell source for further study of new therapy of myocardial disease and building up in vitro model.
