胰岛素样生长因子-1基因转染对兔骨髓间充质干细胞向软骨细胞分化的影响
发布时间：2019-02-15 10:20
【摘要】：目的 关节软骨容易发生损伤和退变,再生能力差,对损伤软骨的实验治疗主要集中在细胞的治疗上,包括应用成熟软骨细胞及其他软骨祖细胞。骨髓间充质干细胞(MSCs)作为成熟软骨细胞的替代品越来越受到重视。MSCs可以很容易从骨髓中获得,增殖传代能力强,细胞均一性好,具多向分化潜能,能分化为各种类型组织,包括软骨和骨。但转移的MSCs在软骨缺损部位还没有产生令人满意的关节软骨。一个可能的问题是局部没有足够能刺激移植细胞分化的细胞因子。体外研究报告显示,特异的蛋白因子能够促进成人MSCs向软骨细胞分化,提高体内软骨修复能力。相关的研究表明,TGF-β1,TGF-β2,TGF-β3,FGF,BMP-2,BMP-6,IGF-1等生长因子具有成软骨能力,因此可以在局部加入这些因子促进MSCs细胞向软骨细胞分化。但外源性细胞因子易于流失、作用时间短、价格昂贵。如果将这些细胞因子基因导入MSCs中,通过自分泌相应细胞因子诱导MSCs向软骨细胞分化,以此基因修饰的MSCs作为种子细胞来构建组织工程软骨,用于软骨缺损和退变治疗具有重要意义。 目前,应用IGF-1基因转染MSCs国内外报道较少。为研究通过IGF-1基因转染来促使MSCs向软骨细胞分化,我们采用密度梯度离心法,与贴壁培养法相结合,对存在于骨髓中的间充质干细胞进行分离、纯化和体外培养扩增,并对其生物学特性进行初步研究。通过构建含有GFP和IGF-1基因的共表达载体,将IGF-1转入MSCs,使MSCs向软骨细胞分化,通过Ⅱ型胶原测定来判断是否稳定表达,以期为开展IGF-1基因治疗软骨缺损的研究奠定基础。 方法 (一)兔骨髓间充质干细胞的分离培养及其生物学活性观察
[Abstract]:Objective the articular cartilage is prone to damage and degeneration, and its regeneration ability is poor. The experimental treatment of damaged cartilage is mainly focused on the cell therapy, including the application of mature chondrocytes and other chondroprogenitor cells. As a substitute for mature chondrocytes, bone marrow mesenchymal stem cells (BMSCs) have attracted more and more attention. MSCs can be easily obtained from bone marrow. Can differentiate into various types of tissue, including cartilage and bone. But the metastatic MSCs did not produce satisfactory articular cartilage at the site of cartilage defect. One possible problem is that there are not enough cytokines locally to stimulate the differentiation of transplanted cells. In vitro studies show that specific protein factors can promote the differentiation of adult MSCs into chondrocytes and improve the ability of cartilage repair in vivo. Related studies have shown that TGF- 尾 1 TGF- 尾 2 and TGF- 尾 3 FGF- 尾 3 FGF- 2 BMP-6 IGF-1 have the ability to form cartilage, so they can be added locally to promote the differentiation of MSCs cells into chondrocytes. But exogenous cytokines are easy to lose, short time and expensive. If these cytokine genes were introduced into MSCs, MSCs was induced to differentiate into chondrocytes by autocrine, and MSCs modified with these genes was used as seed cells to construct tissue engineered cartilage. It is of great significance to treat cartilage defect and degeneration. At present, there are few reports about IGF-1 gene transfection into MSCs at home and abroad. In order to study the differentiation of MSCs into chondrocytes by IGF-1 gene transfection, we used density gradient centrifugation, combined with adherent culture, to isolate, purify and amplify mesenchymal stem cells in bone marrow. And its biological characteristics were preliminarily studied. By constructing a co-expression vector containing GFP and IGF-1 genes, IGF-1 was transferred into MSCs, to differentiate MSCs into chondrocytes, and type 鈪，
本文编号：2423238
[Abstract]:Objective the articular cartilage is prone to damage and degeneration, and its regeneration ability is poor. The experimental treatment of damaged cartilage is mainly focused on the cell therapy, including the application of mature chondrocytes and other chondroprogenitor cells. As a substitute for mature chondrocytes, bone marrow mesenchymal stem cells (BMSCs) have attracted more and more attention. MSCs can be easily obtained from bone marrow. Can differentiate into various types of tissue, including cartilage and bone. But the metastatic MSCs did not produce satisfactory articular cartilage at the site of cartilage defect. One possible problem is that there are not enough cytokines locally to stimulate the differentiation of transplanted cells. In vitro studies show that specific protein factors can promote the differentiation of adult MSCs into chondrocytes and improve the ability of cartilage repair in vivo. Related studies have shown that TGF- 尾 1 TGF- 尾 2 and TGF- 尾 3 FGF- 尾 3 FGF- 2 BMP-6 IGF-1 have the ability to form cartilage, so they can be added locally to promote the differentiation of MSCs cells into chondrocytes. But exogenous cytokines are easy to lose, short time and expensive. If these cytokine genes were introduced into MSCs, MSCs was induced to differentiate into chondrocytes by autocrine, and MSCs modified with these genes was used as seed cells to construct tissue engineered cartilage. It is of great significance to treat cartilage defect and degeneration. At present, there are few reports about IGF-1 gene transfection into MSCs at home and abroad. In order to study the differentiation of MSCs into chondrocytes by IGF-1 gene transfection, we used density gradient centrifugation, combined with adherent culture, to isolate, purify and amplify mesenchymal stem cells in bone marrow. And its biological characteristics were preliminarily studied. By constructing a co-expression vector containing GFP and IGF-1 genes, IGF-1 was transferred into MSCs, to differentiate MSCs into chondrocytes, and type 鈪，
本文编号：2423238
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