CAJ | 학술논문

背景:骨髓间充质干细胞具有多向分化潜能,且可大量体外扩增培养,是重要的组织工程种子细胞.但尚无统一的体外培养及定向诱导方法.目的:探讨体外定向诱导兔骨髓间充质干细胞分化为成骨细胞的可行性.方法:应用密度梯度离心法从兔四肢骨中分离纯化间充质干细胞,应用密度为1.073 g/mL 的 Percol分离液,3000 r/min×30 min 离心,区别于相关报道的 Ficol 分离液,2000-2500 r/min×(20-30) min离心以及全骨髓培养法体外扩增至第3代,分别在普通培养基(对照组)和成骨诱导培养基(实验组)中培养.结果与结论:成功获得大量高纯度骨髓间充质干细胞.经成骨诱导后,实验组骨钙素含量明显高于对照组(P <0.05).实验组碱性磷酸酶和钙结节染色阳性,对照组均阴性.结果表明使用密度梯度离心法可成功建立兔骨髓间充质干细胞的分离培养体系,骨髓间充质干细胞可定向诱导为成骨细胞.
배경:골수간충질간세포구유다향분화잠능,차가대량체외확증배양,시중요적조직공정충자세포.단상무통일적체외배양급정향유도방법.목적:탐토체외정향유도토골수간충질간세포분화위성골세포적가행성.방법:응용밀도제도리심법종토사지골중분리순화간충질간세포,응용밀도위1.073 g/mL 적 Percol분리액,3000 r/min×30 min 리심,구별우상관보도적 Ficol 분리액,2000-2500 r/min×(20-30) min리심이급전골수배양법체외확증지제3대,분별재보통배양기(대조조)화성골유도배양기(실험조)중배양.결과여결론:성공획득대량고순도골수간충질간세포.경성골유도후,실험조골개소함량명현고우대조조(P <0.05).실험조감성린산매화개결절염색양성,대조조균음성.결과표명사용밀도제도리심법가성공건립토골수간충질간세포적분리배양체계,골수간충질간세포가정향유도위성골세포.
@@@@BACKGROUND: Bone marrow mesenchymal stem cells are important seed cells for tissue engineering because of their multi-directional differentiation potential and possibility to be amplified and cultured in vitro. But there have been no uniformed methods of in vitro culture and oriented differentiation. OBJECTIVE: To investigate the feasiblity of in vitro oriented differentation of rabbit bone marrow @@@@mesenchymal stem cells into osteoblasts. METHODS: Rabbit bone marrow mesenchymal stem cells were isolated and purified using density gradient centrifugation (1.073 g/mL Percol separation solution for centrifugation at 3 000 r/min for 30 minutes, which was different from Ficol separation solution for centrifugation at 2 000-2 500 r/min for 20-30 minutes as wel as whole bone marrow culture method). After in vitro amplification, passage 3 bone marrow mesenchymal stem cells were cultured with common culture medium (control group) and osteoblast induction culture medium (experimental group). RESULTS AND CONCLUSION: A large number of high purity bone marrow mesenchymal stem cells were successful y obtained. After osteogenic induction, the content of osteocalcin in the experimental group was significantly higher than that in the control group (P < 0.05). Alkaline phosphatase and calcium tubercle staining were positive in the experimental group, but they were negative in the control group. These findings suggest that density gradient centrifugation can be used to isolate and culture rabbit bone marrow mesenchymal stem cells, and using this method, bone marrow mesenchymal stem cells can be induce-differentiated into osteoblasts.