《中国康复理论与实践》

《中国康复理论与实践》 ›› 2005, Vol. 11 ›› Issue (12): 999-1001.

• 基础研究 • 上一篇    下一篇

两种方法诱导骨髓基质细胞向成骨表型分化的比较

孙崇然1; 景猛2; 李长宇2; 安沂华1; 刘恩重2   

  1. 1.首都医科大学附属北京市神经外科研究所神经干细胞室 北京市 100050;2.哈尔滨医科大学第一临床医学院神经外科 黑龙江哈尔滨市 150001
  • 收稿日期:2005-08-15 出版日期:2005-12-25 发布日期:2005-12-25

Differentiation of rat bone marrow stromal cells into osteogenic phenotype: the comparison of two methods

SUN Chong-ran, JING Meng, LI Chang-yu, et al   

  1. Beijing Neurosurgical Institute of Capital University of Medical Science,100050 Beijing, China
  • Received:2005-08-15 Published:2005-12-25 Online:2005-12-25

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摘要： 目的探讨骨髓基质细胞分化为成骨表型的诱导条件。方法体外扩增大鼠骨髓基质细胞,将第3代细胞分为A组(对照组)、B组(加入地塞米松、β-甘油磷酸钠、维生素C、1,25-二羟维生素D3的诱导组)和C组(在B组基础上加大鼠长骨骨折血肿浸出液诱导组),于诱导后第5、8、11天观察细胞形态变化,用免疫组织化学方法检测诱导细胞的成骨样细胞标志分子(碱性磷酸酶、Ⅰ型胶原、骨钙素),并用von Kossa法检测培养物钙化情况。结果诱导大鼠骨髓基质细胞经小分子诱导培养5d后,部分发生形态学变化,碱性磷酸酶和I型胶原呈弱阳性表达,无钙质沉积;诱导8d后,发生形态学变化的细胞增多,呈集落生长,碱性磷酸酶和I型胶原表达增强,钙质沉积仍不明显;11d后,骨钙素开始表达,且出现钙质沉积。C组细胞表型变化与B组类似,但发生变化的细胞和钙质沉积的数量更多。结论骨髓基质细胞在体外可以被小分子物质诱导为成骨表型;骨折血肿浸出液能够加强这一作用,可作为一种诱导成骨的添加成分。

关键词: 骨髓基质细胞, 诱导分化, 骨折血肿浸出液, 组织工程

Abstract: ObjectiveTo explore environmental conditions under which bone marrow stromal cells could be induced into osteogenic phenotype.MethodsRat bone marrow stromal cells were isolated and proliferated in vitro, and the 3rd passage was divided into the group A (control group), group B (cells cultured in the medium containing dexamethasone, β-glycerol disodium phosphate salt hydrate, vitamin C and active form of vitamin D3), and group C (on the bases of group B, the cells were cultured additionally with fracture hematoma extract). On the post-induction day 5, 8, and 11, the morphological changes were observed and the osteogenic markers such as alkaline phosphotase (ALP), collagen type Ⅰ (Col Ⅰ) and osteocalcin (OCN) were assayed with immunohistochemical staining, the calcification was manifested with von Kossa staining.ResultsIn the group A, no evident osteogenic effects had been observed. In the group B, on 5th day post-induction, some bone marrow stromal cells underwent a morphological change, and mild expression of ALP and Col Ⅰ was observed but with no calcification effect. On 8th day post-induction, the ratio of morphologically changed cells increased, and the expression of ALP and Col Ⅰ increased still with no evident calcification. On 11th day post-induction, anti-OCN staining was positive and the calcium nodes were showed by von Kossa staining. The phenotype changes in the group C were similar to group B, but were more evident.ConclusionBone marrow stromal cells can be induced into osteogenic phenotype in vitro with small molecular inducers. Fracture hematoma extract can enhance this effect thus might be used as an addictive in osteogeneration.

Key words: bone marrow stromal cells, induced differentiation, fracture hematoma extraction, tissue engineering