In vitro chondrogenesis of bone marrow-derived mesenchymal stem cells in a photopolymerizing hydrogel
In vitro chondrogenesis of bone marrow-derived mesenchymal stem cells in a photopolymerizing hydrogel
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DOI：
10.1089/107632703768247377
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发表时间：
2003-08-01
影响因子：
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通讯作者：
Elisseeff, J
中科院分区：
文献类型：
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作者：
Williams, CG;Kim, TK;Elisseeff, J
将来自骨骼成熟山羊的间充质干细胞(MSCs)包裹在聚乙二醇型光聚合水凝胶中，并在有或没有转化生长因子β(1)(TGF)的情况下进行培养，以研究可微创植入的水凝胶支架系统中软骨形成的可能性。通过组织学、生化和RNA分析软骨细胞外基质成分的表达来评价软骨分化。研究的两个对照组分别是单层培养的MSCs和包裹在水凝胶中的MSCs，并在不含转化生长因子-β(1)的软骨细胞培养液中培养6周(6wk-TGF)。包埋细胞的三个实验时间点分别为0天(0d)、3周和6周，在软骨细胞培养液中加入10 ng/ml的转化生长因子-β(1)(3wk+TGF和6wk+TGF)。骨髓间充质干细胞在含有转化生长因子β的水凝胶中增殖(1)。在6wk+TGF构建中，凝胶的糖胺聚糖(GAG)和总胶原含量分别增加到3.5%干重和5.0%干重。免疫组织化学显示有聚集素、连接蛋白和II型胶原的存在。与单层间充质干细胞相比，聚集素和11型胶原基因表达上调。在转化生长因子-β(1)存在的情况下，I型胶原基因的表达在3-6周下降。6wk-TGF水凝胶不产生GAG，仅产生适量的胶原。然而，免疫组织化学和RT-PCR显示该对照组有少量的自发分化。这项研究证明了在体外将MSCs包裹在光聚合水凝胶中形成软骨样组织的能力。该系统可用于微创植入、骨髓间充质干细胞分化和具有多种细胞表型的复合组织结构工程。
Mesenchymal stem cells (MSCs) from skeletally mature goats were encapsulated in a photopolymerizing poly(ethylene glycol)-based hydrogel and cultured with or without transforming growth factor beta(1) (TGF) to study the potential for chondrogenesis in a hydrogel scaffold system amenable to minimally invasive implantation. Chondrogenic differentiation was evaluated by histological, biochemical, and RNA analyses for the expression of cartilage extracellular matrix components. The two control groups studied were MSCs cultured in monolayer and MSCs encapsulated in the hydrogel and cultured for 6 weeks in chondrogenic medium without TGF-beta(1) (6wk-TGF). The three experimental time points for encapsulated cells studied were 0 days (0d), 3 weeks, and 6 weeks in chondrogenic medium with TGF-beta(1) at 10 ng/ml (3wk+TGF and 6wk+TGF). MSCs proliferated in the hydrogels with TGF-beta(1). Glycosaminoglycan (GAG) and total collagen content of the hydrogels increased to 3.5% dry weight and 5.0% dry weight, respectively, in 6wk+TGF constructs. Immunohistochemistry revealed the presence of aggrecan, link protein, and type II collagen. Upregulation of aggrecan and type 11 collagen gene expression compared with monolayer MSCs was demonstrated. Type I collagen gene expression decreased from 3 to 6 weeks in the presence of TGF-beta(1). 6wk-TGF hydrogels produced no GAG and only moderate amounts of collagen. However, immunohistochemistry and RT-PCR demonstrated a small amount of spontaneous differentiation in this control group. This study demonstrates the ability to encapsulate MSCs to form cartilage-like tissue in vitro in a photopolymerizing hydrogel. This system may be useful for minimally invasive implantation, MSC differentiation, and engineering of composite tissue structures with multiple cellular phenotypes.