CAJ | 학술논문

背景：胚胎心脏心外膜细胞是最近被发现的具有分化为心肌细胞和血管平滑肌细胞潜能的心脏干细胞，可分化为心脏三系细胞，为心脏损伤的再生提供了新的细胞来源，但其定向分化机制及调控因素仍不清楚。目的：体外建立小鼠胚胎心脏心外膜细胞培养模型。方法：解剖显微镜下分离11.5-12.5 d小鼠胚胎心脏，剪除肺静脉血管、心房组织及左心室，移至6孔板(或35 mm培养皿)培养，24 h后移走胚胎心脏组织继续培养。相差显微镜观察胚胎心外膜细胞生长特点；使用免疫荧光技术对细胞进行胚胎心外膜细胞特异性抗体Wt-1、Tbx18染色。结果与结论：胚胎心外膜单层细胞从组织块边缘长出，呈鹅卵石样，并围绕组织块向外延伸。移去胚胎心脏后细胞继续生长，且增殖迅速，三四天后长至融合。所有细胞均强阳性表达胚胎心脏心外膜细胞特异性抗体Wt-1及Tbx18。结果表明，胚胎心脏心外膜细胞生长迅速、形态单一，均表达心外膜细胞特异因子，细胞纯度高，成功构建了体外胚胎心脏心外膜细胞培养模型，为研究其定向分化的分子机制提供了新的思路。
배경：배태심장심외막세포시최근피발현적구유분화위심기세포화혈관평활기세포잠능적심장간세포，가분화위심장삼계세포，위심장손상적재생제공료신적세포래원，단기정향분화궤제급조공인소잉불청초。목적：체외건립소서배태심장심외막세포배양모형。방법：해부현미경하분리11.5-12.5 d소서배태심장，전제폐정맥혈관、심방조직급좌심실，이지6공판(혹35 mm배양명)배양，24 h후이주배태심장조직계속배양。상차현미경관찰배태심외막세포생장특점；사용면역형광기술대세포진행배태심외막세포특이성항체Wt-1、Tbx18염색。결과여결론：배태심외막단층세포종조직괴변연장출，정아란석양，병위요조직괴향외연신。이거배태심장후세포계속생장，차증식신속，삼사천후장지융합。소유세포균강양성표체배태심장심외막세포특이성항체Wt-1급Tbx18。결과표명，배태심장심외막세포생장신속、형태단일，균표체심외막세포특이인자，세포순도고，성공구건료체외배태심장심외막세포배양모형，위연구기정향분화적분자궤제제공료신적사로。
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.