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[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。
[Objective] To study the isolation of tendon stem cells from human patellar tendon and to identify the stem cells in vitro. [Method] Muscle stem cells were isolated from human patellar tendon tissue by collagenase digestion and low density inoculation. Through three stages of differentiation and identification, isolated tendon stem cells have the ability to differentiate into osteoblasts, adipocytes and chondrocytes. The expression of the stem cell surface marker factor of the isolated cells was identified by flow cytometry. The expression of tenocyte and chondrocyte related protein was detected by immunohistochemistry. [Result] Muscle stem cells were successfully isolated from human patellar tendon tissue, and about 1.2% ~ 1.4% isolated cells formed single cell clone. The isolated cells express CD44 +, CD105 +, CD29 +, CD45-, and CD14- and have the ability to differentiate into different cells; in addition, tendon stem cells express both tendon-related proteins such as type I collagen and tenascin C Chondrocyte-associated proteins, such as Sox 9 and type II collagen, do not express glycosaminoglycans. [Conclusion] Successful isolation and identification of human patellar stem cells, which have the same stem cell characteristics as mesenchymal stem cells, are different from other mesenchymal stem cells. The successful isolation and identification of tendon stem cells is conducive to the research of later stem cell-related tendon tissue repair, and helps to better understand the function of tendon stem cells in tendon-related diseases and repair.