【摘要】 [目的]研究从人的髌腱组织中分离出肌腱干细胞,并进行体外培养扩增鉴定。[方法]通过胶原酶消化法和低密度接种培养法从人的髌腱组织中分离出肌腱干细胞。通过三期分化鉴定,分离出的肌腱干细胞具有向骨细胞、脂肪细胞和软骨细胞方向分化的能力。通过流式细胞术,鉴定分离出细胞的干细胞表面标记因子的表达。通过免疫组织化学检测其肌腱细胞和软骨细胞相关蛋白的表达。[结果]从人的髌腱组织中成功分离出肌腱干细胞,大约有1.2%~1.4%分离出的细胞形成单细胞克隆。分离出的细胞表达CD44+,CD105+,CD29+,CD45-,和CD14-,并具有向不同细胞分化的能力;另外,肌腱干细胞不仅表达肌腱细胞相关蛋白,如Ⅰ型胶原和tenascin C,也同时会表达软骨细胞相关的蛋白,如Sox 9和Ⅱ型胶原,但其并不表达糖氨多糖。[结论]成功分离鉴定人髌腱干细胞,虽然与间充质干细胞有相同的干细胞特性,是不同于其他间充质干细胞的一种特殊的间充质干细胞。肌腱干细胞的成功分离鉴定有利于以后干细胞相关的肌腱组织修复的研究,并有助于更好的理解肌腱干细胞在肌腱相关疾病和修复的作用。更多还原

【Abstract】 [Objective]This study aimed to isolate and characterize tendon stem cells from human patellar tendon. [Method]Nucleated cells were isolated from human patellar tendon tissues utilizing a collagenase digestion and subsequently plated at a low cell density to allow for the selective proliferation of tendon- derived stem cells. [Result]About 1. 2%- 1. 4% of the cells isolated at low seeding cell density showed clonogenicity. These isolated cells were also found to have osteogenic,chondrogenic,and adipogenic multi- differentiation potential and were CD44+,CD105+,CD29+,CD45-,and CD14-. Furthermore,we identified the expression of tenogenic markers,such as tenascin C and collagen type I. Expression of chondrogenic markers,Sox 9and collagen type Ⅱ,were also observed,but aggrecan was not found. [Conclusion]We successfully isolated tendon- derived stem cells from human patellar tendons. Although they shared some common properties with mesenchymal stem cells,they also exhibited unique characteristics by expressing tenogenic and chondrogenic markers. The successful isolation of tendon- derived stem cells from human patellar tendon will be quite useful for studies on tendon- associated regenerative studies using stem cells,as well as in studies characterizing their roles in tendon tissue- related diseases and healing.更多还原