CAJ | 학술논문

目的建立成年恒河猴骨髓间充质干细胞(rMSC)体外分离、培养及传代扩增的有效体系,为探索新型人工生物骨替代材料提供种子细胞.方法从恒河猴肋骨抽取骨髓,使用淋巴细胞分离液(Ficoll-Hypaque)密度梯度离心法分离骨髓细胞.用含10%胎牛血清(FCS)的低糖DMEM培养基贴壁培养、胰酶消化传代扩增.通过形态学及流式细胞技术对rMSC进行细胞周期分析及表型特征鉴定.结果成年恒河猴骨髓来源的rMSC为成纤维样细胞群体,以梭形的成纤维样细胞为主.流式细胞技术结果显示,rMSC表达CD29、CD44表面分子,不表达CD34、CD45等造血细胞特异性表面分子.结论成年恒河猴骨髓来源的rMSC的分离、培养及传代扩增的细胞群中无造血系细胞污染且符合干细胞增殖特性,为探索新型人工生物骨替代材料的种子细胞提供了一个可靠来源.
목적 건립성년항하후골수간충질간세포(rMSC)체외분리、배양급전대확증적유효체계,위탐색신형인공생물골체대재료제공충자세포.방법 종항하후륵골추취골수,사용림파세포분리액(Ficoll-Hypaque)밀도제도리심법분리골수세포.용함10%태우혈청(FCS)적저당DMEM배양기첩벽배양、이매소화전대확증.통과형태학급류식세포기술대rMSC진행세포주기분석급표형특정감정.결과 성년항하후골수래원적rMSC위성섬유양세포군체,이사형적성섬유양세포위주.류식세포기술결과 현시,rMSC표체CD29、CD44표면분자,불표체CD34、CD45등조혈세포특이성표면분자.결론 성년항하후골수래원적rMSC적분리、배양급전대확증적세포군중무조혈계세포오염차부합간세포증식특성,위탐색신형인공생물골체대재료적충자세포제공료일개가고래원.
Objective To establish method system for isolating and ex-expansion of rhesus marrow mesenchymal stem cells(rMSCs) derived from adult rhesus bone marrow in order to provide seed cells for exploration of new artificial biological bone.Methods Marrow was extracted from healthy adult rhesus rib. Marrow cells were separated by density grade acentric method with Ficoll-Hypaque liquid and then cultured in low glucose DMEM containing 10% fetal cattle serum. After full growth,cells were expended used steapsin. According to morphology and flow cytometer technique,cell cycle and cell-surface antigen profiles of rMSCs were identified.Results rMSCs from adult rhesus bone marrow exhibited fibroblast-like morphology with spindle shape and were positive for antigens of CD29 and CD44,while negative for CD34,CD45 and other haematopoietic cellular specific surface molecular with flow cytometer identification. Conclusion rMSCs derived form adult rhesus bone marrow can be successfully isolated and ex-expanded,with multiplication characteristic of stem cell but without hematopoietic origin,which can provide a reliable source of seed cells for new artificial biological bone research.