转化生长因子-β诱导骨髓基质细胞团体外形成软骨的观察

华西口腔医学杂志

转化生长因子-β诱导骨髓基质细胞团体外形成软骨的观察

陈富林,毛天球,丁桂聪,陈书军,陶凯,顾晓明

710032　第四军医大学口腔医院口腔颌面外科

收稿日期:2003-04-25 修回日期:2003-04-25 出版日期:2003-04-20 发布日期:2003-04-20
基金资助:
本课题为国家“973”项目(编号G1999054308)及国家自然科学基金(编号39970193)资助项目

Experimental Study on the Chondrogenesis Potentiality of Marrow Stromal Cell under the Induction of Transforming Growth Factor-β

CHENFulin,MAO Tianqiu,DINGGuicong,et al·

Oral andMaxillofacialDepartment,College ofStomatology,the FourthMilit- aryMedical University,Xi′an710032,China

Received:2003-04-25 Revised:2003-04-25 Online:2003-04-20 Published:2003-04-20

摘要/Abstract

摘要：

目的　观察骨髓基质细胞(MSCs)在TGF-β诱导下形成软骨的能力,以探讨其作为软骨组织工程种子细胞的可能性。方法　体外分离、培养、扩增兔MSCs,消化后移入离心管中,离心使之成为细胞团,每个细胞团含1×106个MSCs,将细胞团暴露于含20 ng/ml TGF-β的培养液中7 d,然后进行连续培养,观察细胞团的变化,于培养的14 d、28 d取出细胞团,通过体视显微镜、光镜、透射电镜观察软骨的形成情况。结果　TGF-β作用后约14 d,细胞团出现收缩,在离心管底部呈圆球状,以后逐渐增大,28 d后直径最大可达1·8 mm,表面光滑,为亮白色;HE染色显示14 d可见大量活细胞,并有细胞外基质形成,某些局部可见细胞位于陷窝内,28 d可见较多细胞位于陷窝中; 透射电镜观察显示细胞周围有大量胶原基质形成。结论　MSCs在TGF-β作用下可以形成软骨团块,表明其可以向软骨细胞定向分化,能够作为软骨组织工程的种子细胞。

关键词: 组织工程, 软骨, 骨髓基质细胞, 转化生长因子-β

Abstract:

Objective　Seed cell study is an essential area in the research of tissue engineering·To evaluate the potential- ity of marrow stromal cell(MSCs) as seed cell in the regeneration of tissue engineered cartilage, formation of cartilage nodules by culture expanded MSCs pellets under the induction of TGF-βwas investigated.Methods　MSCs were cultured and expandedin vitro. Cell pellets containing 1×106MSCswere obtained by centrifugingMSCs solution at 1 000 r/min in 5 ml centrifugation tube· Pellets were exposed to cell culture media containing 20 ng/mlTGF-βfor 7 days and then cultured for another 7 and 21 days·The nodules were moved out of the tube and cartilage formationwas observed by stereomicroscope, light microscope and electronic mi- croscope·Results　10 days after exposure to TGF-β, pellets contracted and formed small and round nodules on the bottom of the tubes·The nodules grewbigger slowly and reached maximal diameter of 1·8mm in 28 days·The surface of the noduleswas smooth and bright white·Histological examination showed that extra cellular matrix formed in 14 days and in some areas cells situated in lacuna·In 28 days′specimens, a lot of cells situated in lacuna could be observed and the histological appearance looked much similar to cartilage·Electronic microscope observation demonstrated that in 28 days′specimens a large amount of collagen fiber ex- isted·Conclusion　Under the induction of TGF-β, MSCs could differentiate into chondrogenesis cell and form cartilaginous nod- ulesin vitro. This indicated thatMSCs could be the potential seed cells in the regeneration of cartilage employing method of tissue engineering.

Key words: tissue engineering, cartilage, marrow stromal cell, transforming growth factor-β

陈富林,毛天球,丁桂聪,陈书军,陶凯,顾晓明. 转化生长因子-β诱导骨髓基质细胞团体外形成软骨的观察[J]. 华西口腔医学杂志.

CHENFulin,MAO Tianqiu,DINGGuicong,et al·. Experimental Study on the Chondrogenesis Potentiality of Marrow Stromal Cell under the Induction of Transforming Growth Factor-β[J]. West China Journal of Stomatology.

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