Evaluation of a Thermoresponsive Polycaprolactone Scaffold for In Vitro Three-Dimensional Stem Cell Differentiation
Evaluation of a Thermoresponsive Polycaprolactone Scaffold for In Vitro Three-Dimensional Stem Cell Differentiation
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DOI：
10.1089/ten.tea.2013.0710
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发表时间：
2015-01-01
影响因子：
4.1
通讯作者：
Wolbank, Susanne
中科院分区：
文献类型：
--
作者：
Hruschka, Veronika;Saeed, Aram;Wolbank, Susanne
组织工程 (TE) 策略旨在通过使用支持细胞附着、迁移、增殖和分化的生物可吸收支架来模仿自然再生过程。基于将完全可降解聚合物[聚己内酯]与热响应性聚合物（聚乙二醇甲基丙烯酸酯）相结合的想法，开发了一种支架，该支架可在 20 摄氏度以下液化并在 37 摄氏度时固化。在这项研究中，评估了该支架支持 C2C12 细胞和人类脂肪干细胞 (ASC) 的能力，以生成用于软体或骨骼的可扩展三维 (3D) 结构TE。第一步，优化生物材料接种，并对 3D 材料内的细胞附着、存活、分布和持久性进行表征。 C2C12 细胞向成骨和肌源细胞系分化，而 ASC 在对照、脂肪形成或成骨分化培养基中培养。使用定量实时 PCR 检测成骨、肌源和脂肪生成标记物的表达，并通过酶活性和免疫测定来检查分化。两种细胞类型均附着并均匀分布在材料内。 C2C12 细胞和 ASC 表现出在 2D 条件下在所有测试谱系中分化的潜力。在 C2C12 细胞的 3D 成骨条件下，仅骨钙蛋白表达（诱导倍数：16.3 +/- 0.2）和碱性磷酸酶 (ALP) 活性 (p
Tissue engineering (TE) strategies aim at imitating the natural process of regeneration by using bioresorbable scaffolds that support cellular attachment, migration, proliferation, and differentiation. Based on the idea of combining a fully degradable polymer [poly(e-caprolactone)] with a thermoresponsive polymer (polyethylene glycol methacrylate), a scaffold was developed, which liquefies below 20 degrees C and solidifies at 37 degrees C. In this study, this scaffold was evaluated for its ability to support C2C12 cells and human adipose-derived stem cells (ASCs) to generate an expandable three-dimensional (3D) construct for soft or bone TE. As a first step, biomaterial seeding was optimized and cellular attachment, survival, distribution, and persistence within the 3D material were characterized. C2C12 cells were differentiated toward the osteogenic as well as myogenic lineage, while ASCs were cultured in control, adipogenic, or osteogenic differentiation media. Differentiation was examined using quantitative real-time PCR for the expression of osteogenic, myogenic, and adipogenic markers and by enzyme activity and immunoassays. Both cell types attached and were found evenly distributed within the material. C2C12 cells and ASCs demonstrated the potential to differentiate in all tested lineages under 2D conditions. Under 3D osteogenic conditions for C2C12 cells, only osteocalcin expression (fold induction: 16.3 +/- 0.2) and alkaline phosphatase (ALP) activity (p