CAJ | 학술논문

背景:转化生长因子β1是关节软骨组织工程研究的首选生长因子,适当浓度能刺激关节软骨细胞增殖、分裂和分化.目的:建立含有转化生长因子β1的特殊诱导体系培养条件下骨髓间充质干细胞分化为软骨细胞的能力,观察诱导后的细胞形态及表型变化.方法:于兔胫骨结节内侧抽取骨髓,采用贴壁培养法分离纯化兔骨髓间充质干细胞,取第3代骨髓间充质干细胞行流式细胞仪检测鉴定其表面抗原,以含有转化生长因子β1的特殊软骨诱导体系的培养条件下对第3代骨髓间充质干细胞诱导培养21 d,诱导后与人鼻中隔软骨细胞进行比较.采用免疫组织化学法对Ⅱ型胶原进行定性检测.结果与结论:贴壁培养法可分离并纯化兔骨髓间充质干细胞,所得第3代骨髓间充质干细胞表面抗原CD44 阳性,CD34、CD45 阴性.经诱导培养21 d后细胞形态变为不规则,Ⅱ型胶原免疫组织化学染色显示可见阳性细胞.提示含有转化生长因子β1的特殊软骨诱导体系的培养条件下,骨髓间充质干细胞可以转化为软骨细胞,且与正常软骨细胞无明显差异.
배경:전화생장인자β1시관절연골조직공정연구적수선생장인자,괄당농도능자격관절연골세포증식、분렬화분화.목적:건립함유전화생장인자β1적특수유도체계배양조건하골수간충질간세포분화위연골세포적능력,관찰유도후적세포형태급표형변화.방법:우토경골결절내측추취골수,채용첩벽배양법분리순화토골수간충질간세포,취제3대골수간충질간세포행류식세포의검측감정기표면항원,이함유전화생장인자β1적특수연골유도체계적배양조건하대제3대골수간충질간세포유도배양21 d,유도후여인비중격연골세포진행비교.채용면역조직화학법대Ⅱ형효원진행정성검측.결과여결론:첩벽배양법가분리병순화토골수간충질간세포,소득제3대골수간충질간세포표면항원CD44 양성,CD34、CD45 음성.경유도배양21 d후세포형태변위불규칙,Ⅱ형효원면역조직화학염색현시가견양성세포.제시함유전화생장인자β1적특수연골유도체계적배양조건하,골수간충질간세포가이전화위연골세포,차여정상연골세포무명현차이.
BACKGROUND: Transforming growth factor beta 1 (TGF-β1) is a first selected growth factor in study of articular cartilage tissue engineering. Suitable concentration can stimulate proliferation, division and differentiation of articular chondrocytes. OBJECTIVE: To establish an special inducing system for differentiation of bone marrow mesenchymal stem cells (BMSCs) into chondrocytes in contained TGF-β1 culture in vivo,and to observe the changes in cell form and phenotype.METHODS: BMSCs were separated and purified from rabbit tibial tubercle using attachment culture method. Surface antigen of the third-generated BMSCs was determined by using flow cytometry. Subsequently,the third-generated BMSCs were induced with TGF-β1-contained special inducing system in 21-day culture, which was compared with human septal cartilage cells of nose following induction. Collagen type Ⅱ was qualitatively determined using immunohistochemical method.RESULTS AND CONCLUSION: Rabbit BMSCs were separated and purified using attachment culture method. The third-generated BMSCs were positive for surface antigen CD44,but negative for surface antigen CD34 and CD45. Cell form was irregular 21 days after induction. Immunohistochemical staining for type II collagen demonstrated positive cells. Results suggested that under culture system containing TGF-β1 BMSCs may directionally differentiate into chondrocytes,and there were not significant differences with normal chondrocytes.