Establishment of in vitro culture model of mouse embryonic heart epicardial cells
10.3969/j.issn.2095-4344.2013.44.001
- VernacularTitle:小鼠胚胎心脏心外膜细胞体外培养模型的建立
- Author:
Bin ZENG
;
Aili WANG
;
Dongdong HU
;
Chang LI
- Publication Type:Journal Article
- Keywords:
embryonic stem cells;
cell culture techniques;
regeneration;
cell proliferation
- From:
Chinese Journal of Tissue Engineering Research
2013;(44):7661-7666
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:The embryonic epicardium can differentiate into myocardial cells and the cardiac stem cells with the potential of vascular smooth muscle cells, and it can differentiate into cardiac three-line cells which provide a new cellsource for the regeneration of cardiac injury. But the directed differentiation mechanisms and regulatory factors are stil unclear. OBJECTIVE:To establish the in vitro culture model of epicardial cells of mouse embryonic heart. METHODS:Embryonic hearts were dissected from the mice at pregnant 10.5-11.5 days, and the pulmonary veins, atrial tissue and left ventricle were cut off, then the embryonic hearts were transplanted into the 6-wel plates (or 35 mm dish) for culture. After cultured for 24 hours, the embryonic heart tissues were removed and cultured continuously. Phase contrast microscope was used to observe the growth characteristics of embryonic epicardial cells;the immunofluorescence technique was used to stain the specific antibody Wt-1 and Tbx18 of embryonic epicardial cells. RESULTS AND CONCLUSION:The embryonic epicardial cellmonolayers grew from tissue block edge with cobblestone-like and extended outwardly around the tissue blocks. After removed the embryonic heart, the cells grew continuously with rapid proliferation, and got fusion at 34 days. Al the embryonic epicardial cells could positively express the specific antibody Wt-1 and Tbx18 of embryonic epicardial cells. The results indicate that embryonic epicardial cells have the characteristics of rapidly growth and uniform morphology, and can express the embryonic epicardial cellspecific antibody with high purity. The successful y constructed in vitro culture models of embryonic epicardial cells provide new idea for the molecular mechanisms of directed differentiation.
