CAJ | 학술논문

目的：对4种建立滋养细胞的方法进行比较，以期寻求一种简便高效的人早孕滋养细胞的体外培养方法，为相关研究提供基础。方法早孕绒毛通过胰酶联合胶原酶消化分离后，分别采用直接接种、差速贴壁联合差速消化法、人外周血淋巴细胞分离液分离纯化法、完整绒毛消化联合简化的 percoll 密度梯度分离法4种方法纯化培养，倒置显微镜观察细胞形态，应用 HE 染色、免疫组化和免疫细胞化学法检测细胞纯度。结果直接接种、差速贴壁联合差速消化法、人外周血淋巴细胞分离液分离纯化法得到的细胞细胞角蛋白7（CK －7）表达阳性率不足60％；简化的 percoll 密度梯度分离法得到的细胞 CK －7表达阳性率达90％以上。结论完整绒毛消化联合简化的 percoll 密度梯度分离纯化可以得到大量较高纯度的滋养细胞以供后期实验。
목적：대4충건립자양세포적방법진행비교，이기심구일충간편고효적인조잉자양세포적체외배양방법，위상관연구제공기출。방법조잉융모통과이매연합효원매소화분리후，분별채용직접접충、차속첩벽연합차속소화법、인외주혈림파세포분리액분리순화법、완정융모소화연합간화적 percoll 밀도제도분리법4충방법순화배양，도치현미경관찰세포형태，응용 HE 염색、면역조화화면역세포화학법검측세포순도。결과직접접충、차속첩벽연합차속소화법、인외주혈림파세포분리액분리순화법득도적세포세포각단백7（CK －7）표체양성솔불족60％；간화적 percoll 밀도제도분리법득도적세포 CK －7표체양성솔체90％이상。결론완정융모소화연합간화적 percoll 밀도제도분리순화가이득도대량교고순도적자양세포이공후기실험。
Objective To study the methodology for purification of human first trimester trophoblast cells from placentas with different methods , thus to seek a simple method for purification of human first trimester trophoblast cells in vitro.Methods Target cells isolated from first trimester human placenta with trypsin and collagenase Ⅰ were incubated directly; purified by differential time attachment and digestion ; purified by lymphocyte separation medium ; and dissociated in trypsin /collagenase Ⅰ with purification by simplified percoll gradient centrifugation , respectively.Morphologic observa-tion was applied with inverted phase contrast microscope , with purity was assessed by HE staining and immunocytochemis -try.Results Cytokeratin -7 positive cells were more than 90% only with the last method, while cytokeratin -7 positive cells were less than 60% with the other methods.Conclusion First trimester human trophoblast cells can be obtained through whole first trimester human placenta digestion and followed by simplified percoll gradient centrifugation purification .