CAJ | 학술논문

背景:分离培养鼠、兔骨髓间充质干细胞的科研实践较多,而组织工程临床实践大多以同种属种子细胞的科研为基础.目的:建立一种稳定、高效,满足临床和实验室需要的人骨髓间充质干细胞分离培养方法. 方法:使用骨穿针于人髂前上棘处抽取骨髓和松质骨,用15 mL 培养体系冲洗松质骨并收集冲洗液,采用直接培养法,利用细胞贴壁性能初筛细胞;流式细胞仪检测细胞表面抗原(CD29、CD34、CD44、CD45、CD105),分别进行成骨、成脂肪诱导分化,在诱导过程中进行细胞形态学观察对干细胞进行初步鉴定. 结果与结论:松质骨冲洗液直接培养法在培养第5天的时候出现细胞集落,细胞稳定表达 CD29、CD44、CD105,不表达 CD34、CD45.成骨诱导20 d 后出现明显钙结节,茜素红染色呈红色结节.成脂诱导7 d 后脂滴明显形成,油红素 O 染色见大量脂质沉淀.结果可见采用松质骨冲洗液直接培养法可以从人松质骨中短时间内获得大量间充质干细胞,其具有良好的细胞形态,稳定表达的细胞抗原及成骨、成脂分化能力.
배경:분리배양서、토골수간충질간세포적과연실천교다,이조직공정림상실천대다이동충속충자세포적과연위기출.목적:건립일충은정、고효,만족림상화실험실수요적인골수간충질간세포분리배양방법. 방법:사용골천침우인가전상극처추취골수화송질골,용15 mL 배양체계충세송질골병수집충세액,채용직접배양법,이용세포첩벽성능초사세포;류식세포의검측세포표면항원(CD29、CD34、CD44、CD45、CD105),분별진행성골、성지방유도분화,재유도과정중진행세포형태학관찰대간세포진행초보감정. 결과여결론:송질골충세액직접배양법재배양제5천적시후출현세포집락,세포은정표체 CD29、CD44、CD105,불표체 CD34、CD45.성골유도20 d 후출현명현개결절,천소홍염색정홍색결절.성지유도7 d 후지적명현형성,유홍소 O 염색견대량지질침정.결과가견채용송질골충세액직접배양법가이종인송질골중단시간내획득대량간충질간세포,기구유량호적세포형태,은정표체적세포항원급성골、성지분화능력.
BACKGROUND: Experiments for the isolation and culture of rat or rabbit bone marrow mesenchymal stem cel s are more common, but the clinical experiments of tissue engineering are often on the basis of the seed cel s of the same species. @@@@OBJECTIVE: To establish a protocol for isolating and culturing human bone marrow mesenchymal stem cel s which is stable and efficient and consistent with the needs of clinical and laboratory experiments. @@@@METHODS: Bone marrow and spongy bone were obtained from the human anterior superior iliac spine, and then the bone marrow, the spongy bone was flushed by 15 mL culture medium and then the rinse solution was col ected. The cel s were primary screened on the basis of adhesion function by direct cultivation method; the surface antigens, including CD29, CD34, CD44, CD45 and CD105 were detected by flow cytometry. Bone marrow mesenchymal stem cel s were differentiated into osteoblasts and adipocytes, and the differentiated mesenchymal stem cel s were identified by morphological observation. @@@@RESULTS AND CONCLUSION: Cel colony could be seen at 5 days after cultured with spongy bone washing liquid. These cel s were uniformly negative for CD34, CD45 and positive for CD29, CD44 and CD105. Calcium nodules were observed after osteogenic induction for 20 days and positive for alizarin red staining. The lipid droplets could be seen after adipogenic induction for 7 days and oil red O staining showed a large number of lipid deposition. The study shows that a large amount of mesenchymal stem cel s can be isolated and cultured from adult human spongy bone in short time by direct cultivation methods, and the mesenchymal stem cel s are uniform in morphology, the cel s can express the antigens stably and differentiate into osteoblasts and adipocytes.