CAJ | 학술논문

目的：探讨树鼩骨髓间充质干细胞（ BM-MSCs）的体外分离、传代及定向诱导为脂肪细和成骨细胞的可行性。方法通过密度梯度离心联合贴壁培养法对树鼩骨髓间充质干细胞进行体外分离、扩增、纯化，倒置相差显微镜进行形态学观察。用成脂诱导液（ DMEM/F12＋10％FBS＋100 U/mL青霉素＋100μg/mL链霉素＋1.0μmol/L地塞米松＋0.2 mmol/L吲哚美辛＋0.01 mg/mL胰岛素＋0.5 mmol/L IBMX）和成骨诱导液（高糖DMEM＋10％FBS＋100 U/mL青霉素＋100μg/mL链霉素＋50 ng/mL BMP-2）对分离的树鼩BM-MSCs分别定向诱导为脂肪细胞和成骨细胞。结果原代和传代细胞为梭形或三角形，可增殖形成克隆。 BM-MSCs成脂诱导后油红O染色细胞内出现红色脂滴，成骨诱导后茜素红染色可观察到矿化结节。结论密度梯度离心联合贴壁培养法分离培养树鼩BM-MSCs简便可行，获得的BM-MSCs可体外诱导分化为脂肪细胞和成骨细胞。
목적：탐토수구골수간충질간세포（ BM-MSCs）적체외분리、전대급정향유도위지방세화성골세포적가행성。방법통과밀도제도리심연합첩벽배양법대수구골수간충질간세포진행체외분리、확증、순화，도치상차현미경진행형태학관찰。용성지유도액（ DMEM/F12＋10％FBS＋100 U/mL청매소＋100μg/mL련매소＋1.0μmol/L지새미송＋0.2 mmol/L신타미신＋0.01 mg/mL이도소＋0.5 mmol/L IBMX）화성골유도액（고당DMEM＋10％FBS＋100 U/mL청매소＋100μg/mL련매소＋50 ng/mL BMP-2）대분리적수구BM-MSCs분별정향유도위지방세포화성골세포。결과원대화전대세포위사형혹삼각형，가증식형성극륭。 BM-MSCs성지유도후유홍O염색세포내출현홍색지적，성골유도후천소홍염색가관찰도광화결절。결론밀도제도리심연합첩벽배양법분리배양수구BM-MSCs간편가행，획득적BM-MSCs가체외유도분화위지방세포화성골세포。
Objective To study the isolation,culture, adipogenic and osteogenic induction Tupaia bone marrow mesenchymal stem cells(BM-MSCs).Method The BM-MSCs from tupaia were isolated and expended by combination of gradient centrifugation and adherence culture , then subcultured and observed for morphology under inverted phase contrast microscope.BM-MSCs were induced to adipocytes .and osteoblasts in vitro Result Cells were spindle or triangle-shaped, and clone proliferation .Cells were successfully induced into adipocytes .and osteoblasts Conclusions The method of isolation BM-MSCs from tupaia by combination of gradient centrifugation and adherence culture is simple and feasible , BM-MSCs have differentiation potential into adipocytes and osteoblasts .