Comparison of Mesenchymal Stem Cell Source Differentiation Toward Human Pediatric Aortic Valve Interstitial Cells within 3D Engineered Matrices
Comparison of Mesenchymal Stem Cell Source Differentiation Toward Human Pediatric Aortic Valve Interstitial Cells within 3D Engineered Matrices
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DOI：
10.1089/ten.tec.2014.0589
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发表时间：
2015-08-01
影响因子：
3
通讯作者：
Butcher, Jonathan T.
中科院分区：
文献类型：
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作者：
Duan, Bin;Hockaday, Laura A.;Butcher, Jonathan T.
对于需要具有生长和生物整合能力的替代物的儿童来说，活的组织工程心脏瓣膜(TEHV)将是一个主要的好处。TEHV面临的一个长期挑战是可获得的人类细胞来源(S)，它可以模拟对长期功能至关重要的天然瓣膜细胞表型和基质重塑特征。来源于骨髓或脂肪组织的间充质干细胞(BMMSC)或脂肪组织间充质干细胞(ADMSC)是TEHV的诱人细胞来源，但尚未在相关的三维环境中与儿童人主动脉瓣间质细胞(PHAVIC)进行比较。在这项研究中，我们比较了ADMSC和BMMSC与PHAVIC在材料模数与主动脉瓣瓣相似的三维生物活性杂化水凝胶中的自发和诱导多能性。PHAVIC对诱导培养基具有一定的多系分化能力，但仅限于早期阶段，远低于ADMSC和BMMSC。在含有碱性成纤维细胞生长因子的HAVIC培养液中，ADMSC表达的细胞表型标志更接近于PHAVIC，而BMMSC表现出与PHAVIC相似的细胞外基质重塑特征。最后，我们将碱性成纤维细胞生长因子共价连接到聚乙二醇单丙烯酸酯连接物上，并进一步共价固定在三维杂化水凝胶中。固定化碱性成纤维细胞生长因子上调Vimentin表达，促进PHAVIC、ADMSC和BMMSC向成纤维细胞分化。这些发现表明，干细胞在3D培养中保持了较高的成骨分化能力，但可以通过bFGF的基质连接向成纤维细胞分化。这样的策略对于在心脏瓣膜组织工程应用中利用干细胞来源可能是重要的。
Living tissue-engineered heart valves (TEHV) would be a major benefit for children who require a replacement with the capacity for growth and biological integration. A persistent challenge for TEHV is accessible human cell source(s) that can mimic native valve cell phenotypes and matrix remodeling characteristics that are essential for long-term function. Mesenchymal stem cells derived from bone marrow (BMMSC) or adipose tissue (ADMSC) are intriguing cell sources for TEHV, but they have not been compared with pediatric human aortic valve interstitial cells (pHAVIC) in relevant 3D environments. In this study, we compared the spontaneous and induced multipotency of ADMSC and BMMSC with that of pHAVIC using different induction media within three-dimensional (3D) bioactive hybrid hydrogels with material modulus comparable to that of aortic heart valve leaflets. pHAVIC possessed some multi-lineage differentiation capacity in response to induction media, but limited to the earliest stages and much less potent than either ADMSC or BMMSC. ADMSC expressed cell phenotype markers more similar to pHAVIC when conditioned in basic fibroblast growth factor (bFGF) containing HAVIC growth medium, while BMMSC generally expressed similar extracellular matrix remodeling characteristics to pHAVIC. Finally, we covalently attached bFGF to PEG monoacrylate linkers and further covalently immobilized in the 3D hybrid hydrogels. Immobilized bFGF upregulated vimentin expression and promoted the fibroblastic differentiation of pHAVIC, ADMSC, and BMMSC. These findings suggest that stem cells retain a heightened capacity for osteogenic differentiation in 3D culture, but can be shifted toward fibroblast differentiation through matrix tethering of bFGF. Such a strategy is likely important for utilizing stem cell sources in heart valve tissue engineering applications.