犬骨髓间充质干细胞向瓣膜间质细胞诱导分化的实验研究

发布时间：2018-02-26 21:36

本文关键词： 组织工程心脏瓣膜去细胞瓣膜骨髓间充质干细胞诱导分化间质细胞　出处：《第四军医大学》2005年硕士论文　论文类型：学位论文

【摘要】：瓣膜置换术是治疗瓣膜疾病的有效方法,但现有的人工心脏瓣膜都有不容忽视的缺点:机械瓣需要终生抗凝;生物瓣膜易钙化衰败有的仍需抗凝;同种异体瓣膜来源有限。组织工程心脏瓣膜(TEHV)按照组织工程学方法,利用可降解材料构建瓣膜支架,种植的自体细胞在支架上生长并产生细胞外基质(ECM),逐渐形成活的瓣膜组织。它有良好的组织相容性,可避免血栓栓塞及钙化衰败等问题,有自我修复能力,随机体生长,因此被认为是一种理想的瓣膜替代物。种子细胞是组织工程瓣膜的上游问题,血管间质细胞因与瓣膜间质细胞同处于血流环境而首先被考虑作为组织工程瓣膜间质的种子细胞,但是其原代培养相对繁琐且要牺牲一根完整血管,为有创性,和此相比,骨髓间充质干细胞获取相对容易,创伤小、易扩增、无免疫原性且具多分化潜能而引起关注。在本研究中,我们就骨髓间充质干细胞诱导分化,并将诱导后骨髓间充质细胞与血管间质细胞作为组织工程瓣膜种子细胞进行了对比研究工作。本研究分为三部分: 第一部分:用Percoll密度梯度离心法分离骨髓单个核细胞并经贴壁培养纯化而得犬骨髓间充质干细胞(BMSCs)、对其进行鉴定、观察其生物学特性,并对犬BMSCs进行诱导分化,对诱导而来的细胞,同样也观察了其生物学性能,然后作进一步的鉴定。并对诱导细胞进行冻存复苏,观察其复苏率。结果:经Percoll液分离的骨髓单核细胞,经贴壁
[Abstract]:Valvular replacement is an effective method for the treatment of valvular diseases, but the existing artificial heart valves have some shortcomings that can not be ignored: mechanical valves need lifetime anticoagulant, biological valves are prone to calcification decay and still need anticoagulant. The tissue engineering heart valve (TEHVV) uses biodegradable materials to construct the valve scaffold according to the tissue engineering method. The implanted autologous cells grow on the scaffold and produce extracellular matrix (ECM), which gradually form live valve tissue. It has good histocompatibility, can avoid thromboembolism and calcification decay, and has the ability to repair itself and grow at random. Seed cells are the upstream problem of tissue engineering valves. Vascular interstitial cells are considered as seed cells of tissue engineering valves because they are in the same blood flow environment as valve interstitial cells. But its primary culture is relatively cumbersome and requires the sacrifice of a complete blood vessel to be invasive. Compared with this, bone marrow mesenchymal stem cells are relatively easy to obtain, have less trauma, and are easy to expand. In this study, we induced differentiation of bone marrow mesenchymal stem cells (BMSCs). The mesenchymal cells and vascular mesenchymal cells were used as seed cells of tissue engineering valve. The first part: bone marrow mononuclear cells were isolated by Percoll density gradient centrifugation and purified by adherent culture. BMSCs were identified, their biological characteristics were observed, and canine BMSCs was induced to differentiate. The biological properties of induced cells were also observed and further identified. The induced cells were resuscitated by cryopreservation and the rate of resuscitation was observed. Results: bone marrow monocytes isolated from Percoll solution were adherent to the wall.
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R329.2

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