CAJ | 학술논문

目的：脐带血间充质干细胞为组织工程种子细胞领域研究热点，但目前仍未得到安全、高效的脐带血间充质干细胞分离培养体系，该实验研究旨在探讨脐带血间充质干细胞的适宜培养条件。方法无菌条件下取足月正常新生儿脐带血，随机分为高糖DMEM组、低糖DMEM组以及脐带血间充质干细胞组。分离提取的脐带血单个核细胞分别以1×104、1×105和1×106个细胞／ml的接种密度接种于脐带血间充质干细胞培养基中，观察3种培养基及不同接种密度培养下的细胞贴壁生长情况，并用流式细胞技术分析细胞的表面抗原。结果高糖DMEM组未见到贴壁生长的间充质干细胞，低糖DMEM组及脐带血间充质干细胞组均可见到贴壁生长的梭形成纤维样细胞（P＜0.05），在不同的接种密度条件下，1×105组及1×106组接种密度可见到成纤维样贴壁细胞，1×105组得到的细胞数量及细胞生长情况更好，1×104组未见到细胞贴壁生长。结论分离提取得到的脐带血单个核细胞在T 25培养瓶中最佳接种密度为1×105／ml，最适合培养基为脐带血间充质干细胞培养基。
목적：제대혈간충질간세포위조직공정충자세포영역연구열점，단목전잉미득도안전、고효적제대혈간충질간세포분리배양체계，해실험연구지재탐토제대혈간충질간세포적괄의배양조건。방법무균조건하취족월정상신생인제대혈，수궤분위고당DMEM조、저당DMEM조이급제대혈간충질간세포조。분리제취적제대혈단개핵세포분별이1×104、1×105화1×106개세포／ml적접충밀도접충우제대혈간충질간세포배양기중，관찰3충배양기급불동접충밀도배양하적세포첩벽생장정황，병용류식세포기술분석세포적표면항원。결과고당DMEM조미견도첩벽생장적간충질간세포，저당DMEM조급제대혈간충질간세포조균가견도첩벽생장적사형성섬유양세포（P＜0.05），재불동적접충밀도조건하，1×105조급1×106조접충밀도가견도성섬유양첩벽세포，1×105조득도적세포수량급세포생장정황경호，1×104조미견도세포첩벽생장。결론분리제취득도적제대혈단개핵세포재T 25배양병중최가접충밀도위1×105／ml，최괄합배양기위제대혈간충질간세포배양기。
Objective To explore the appropriate culture conditions of umbilical cord blood mesenchymal stem cells.Methods The sterile cord blood from normal full-term newborns was randomly divided into three groups:the high sugar DMEM group , the low sugar DMEM group and the umbilical cord blood mesenchymal stem cells be -tween groups .Extractions of umbilical cord blood mononuclear cells were inoculated in umbilical cord blood mesen -chymal stem cells in the culture media by the density of 1 ×105 , 1 ×104 , and 1 ×106 cells/ml respectively.The three kinds of media and different inoculation density culture adherent cell growth situations were observed and analy -sis of cell surface antigen was performed using flow cytometry technique .Results Ectomesenchymal stem cells of sugar DMEM group did not grow on the sidewall , while the growth of the spindle formation fiber sample cells was found on the sidewall in low sugar DMEM group and umbilical cord blood mesenchymal stem cell ( P<0.05 ) .Of dif-ferent inoculation densities , cells grew best at the density of 1 ×105 , followed by 1 ×106 .Cells failed to grow at the density of 1 ×104 .Conclusion Umbilical cord blood mononuclear cells in T 25 culture bottle grow best at the inocu-lation density of 1 ×105/ml, which is the most suitable culture medium for umbilical cord blood mesenchymal stem cell.