CAJ | 학술논문

目的：探讨建立一种新的从膨胀液中提取脂肪间充质干细胞（ADSCs）的分离方法。方法：收集含有脂肪间充质干细胞的膨胀液，然后从膨胀液中分离出脂肪间充质干细胞并进行体外培养，观察培养间充质干细胞生长状态，流式细胞术检测间充质干细胞干性标记物，细胞生长曲线比较新方法与运用传统方法分离脂肪间充质干细胞的增殖活性，多向诱导分化鉴定其向成骨，成软骨及成脂方向分化的能力。结果：成功建立了一种新的从膨胀液中提取脂肪间充质干细胞的分离方法；分离自膨胀液的间充质干细胞数量虽然低于与等体积脂肪组织来源的间充质干细胞，但细胞生长曲线分析结果表明其增殖速度快，生长至第8天时，密度基本等同于脂肪组织来源间充质干细胞。间充质干细胞表面分子标记物CD73，CD90，CD105，CD45，CD34，CD11b，CD19，HLA—DR表达测定结果显示正常，阳性细胞率与脂肪组织来源的干细胞相近。多向诱导分化结果显示从膨胀液中分离的脂肪间充质干细胞可以向成脂、成骨和成软骨三向分化。结论：新方法分离的细胞确为脂肪间充质干细胞，符合国际干细胞协会规定的定义标准。
목적：탐토건립일충신적종팽창액중제취지방간충질간세포（ADSCs）적분리방법。방법：수집함유지방간충질간세포적팽창액，연후종팽창액중분리출지방간충질간세포병진행체외배양，관찰배양간충질간세포생장상태，류식세포술검측간충질간세포간성표기물，세포생장곡선비교신방법여운용전통방법분리지방간충질간세포적증식활성，다향유도분화감정기향성골，성연골급성지방향분화적능력。결과：성공건립료일충신적종팽창액중제취지방간충질간세포적분리방법；분리자팽창액적간충질간세포수량수연저우여등체적지방조직래원적간충질간세포，단세포생장곡선분석결과표명기증식속도쾌，생장지제8천시，밀도기본등동우지방조직래원간충질간세포。간충질간세포표면분자표기물CD73，CD90，CD105，CD45，CD34，CD11b，CD19，HLA—DR표체측정결과현시정상，양성세포솔여지방조직래원적간세포상근。다향유도분화결과현시종팽창액중분리적지방간충질간세포가이향성지、성골화성연골삼향분화。결론：신방법분리적세포학위지방간충질간세포，부합국제간세포협회규정적정의표준。
Purpose： To explore and establish a new method of isolating adipose-derived stem cell （ADSCs） from tumescent liquid. Methods： Human tumescent liquid containing adipose-derived stem cell was collected and adipose-derived stem cells were isolated and expanding cultured in vitro. The growth and proliferation activity of adipose-derived stem cell were systematically analyzed. Moreover, growth curves of cells isolated with different approach were drawn and surface antigens against mesenchymal stem cell markers were detected with flow cytometry. Muhi-direetional differentiation properties including adipocytic trans-differentiation, osteogenic differentiation and chondrogenic differentiation of ADSCs had been verified. Results： The method of isolating adipose-derived stem cell from tumescent liquid was established. The cell number from this method was lower when compared to the old method that directly isolate adipose-derived stem ceils from the same volume of adipose tissue, however, growth curves analysis demonstrated that stem cells isolated from new method increased quickly. Similar to stem cells from adipose tissues, these stem cells could achieve plateau phase at 8 day. Meanwhile, adipose-derived stem cell surface markers, including CD73, CD90, CDI05, CD45, CD34, CD1 lb, CD19, HLA-DR were normally expressed and positive stem cells were identical in two methods which were detected by flow cytometry. Furthermore, adipose-derived stem cells were identified with multi-directional differentiation properties including adipocytic trans-differentiation, osteogenic differentiation and chondrogenic differentiation. Conclusion： cells isolated with new method had been verified as real adipose-derived stem cells and were in accord with the defination standard of international society for stem cell research.