CAJ | 학술논문

背景：研究发现充足的血供是骨组织修复与再生必不可少的条件。 　　目的：应用血管内皮祖细胞与骨髓间充质干细胞构建出复合细胞膜片。 　　方法：体外分离培养SD大鼠血管内皮祖细胞及骨髓间充质干细胞，将两种细胞直接共培养，经成膜诱导10 d后构建出复合细胞膜片并对获得的膜片进行大体、倒置显微镜及苏木精-伊红染色观察。将血管内皮祖细胞及骨髓间充质干细胞分别应用荧光标记液标记后，观察膜片诱导过程中两种细胞的分布及交流情况。对复合细胞膜片进行碱性磷酸酶活性检测及茜素红染色，观察其成骨分化能力。 　　结果与结论：血管内皮祖细胞与骨髓间充质干细胞直接共培养成膜诱导10 d后可成功获得复合细胞膜片，两种细胞在成膜诱导过程中存在细胞间接触。获得的膜片由多层细胞及细胞外基质构成，经碱性磷酸酶活性检测及茜素红染色结果证实该膜片具有较强的成骨分化能力，为进一步将血管内皮祖细胞/骨髓间充质干细胞复合细胞膜片应用于骨缺损的修复提供了条件。
배경：연구발현충족적혈공시골조직수복여재생필불가소적조건。 　　목적：응용혈관내피조세포여골수간충질간세포구건출복합세포막편。 　　방법：체외분리배양SD대서혈관내피조세포급골수간충질간세포，장량충세포직접공배양，경성막유도10 d후구건출복합세포막편병대획득적막편진행대체、도치현미경급소목정-이홍염색관찰。장혈관내피조세포급골수간충질간세포분별응용형광표기액표기후，관찰막편유도과정중량충세포적분포급교류정황。대복합세포막편진행감성린산매활성검측급천소홍염색，관찰기성골분화능력。 　　결과여결론：혈관내피조세포여골수간충질간세포직접공배양성막유도10 d후가성공획득복합세포막편，량충세포재성막유도과정중존재세포간접촉。획득적막편유다층세포급세포외기질구성，경감성린산매활성검측급천소홍염색결과증실해막편구유교강적성골분화능력，위진일보장혈관내피조세포/골수간충질간세포복합세포막편응용우골결손적수복제공료조건。
BACKGROUND:Many studies have showed that enough blood supply is an essential condition of bone repair and regeneration. OBJECTIVE:To construct the endothelial progenitor cells/bone marrow mesenchymal stem cells (EPCs/BMSCs) composite sheet. METHODS:After isolation and culture, EPCs and BMSCs were co-cultured directly to form EPCs/BMSCs sheet by cellsheet-inducing medium. After 10 days of induction, the sheet was investigated by gross observation, inverted microscope and hematoxylin-eosin staining. The distribution and communication of EPCs and BMSCs during the process of cellsheet induction were observed after the fluorescence labeling separately. Alkaline phosphatase assay and alizarin red staining were applied to examine the ability of osteogenic differentiation of EPCs/BMSCs sheet. RESULTS AND CONCLUSION:EPCs/BMSCs sheet was harvested after 10-day induction. Cel-cellcontact between EPCs and BMSCs could be observed during the process of the cellsheet preparation. The harvested sheet was composed of multiple layers of cells and cel-produced extracellular matrix. Alkaline phosphatase assay and alizarin red staining both demonstrated that EPCs/BMSCs sheet had good osteogenic differentiation ability. These results suggested that EPCs/BMSCs sheet can be constructed successful y, and the sheet has strong osteogenic differentiation capability in vitro, providing the foundation for the repair of bone defects.