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首页> 外文期刊>Tissue engineering, Part A >An Injectable Recombinant Collagen I Peptide-Based Macroporous Microcarrier Allows Superior Expansion of C2C12 and Human Bone Marrow-Derived Mesenchymal Stromal Cells and Supports Deposition of Mineralized Matrix
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An Injectable Recombinant Collagen I Peptide-Based Macroporous Microcarrier Allows Superior Expansion of C2C12 and Human Bone Marrow-Derived Mesenchymal Stromal Cells and Supports Deposition of Mineralized Matrix

机译：可注射的重组胶原I肽基大孔微载体允许C2C12和人骨髓衍生的间充质细胞的优异膨胀，并支持矿化基质的沉积

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The development of scaffold formulations based on extracellular matrix (ECM)-inspired synthetic materials constitutes an important resource for the advance of cell-based therapies in bone tissue engineering approaches, where both cell and scaffold implantation are often needed. Culturing cells on porous microcarriers (MCs) allows cell expansion in a three-dimensional microenvironment and constitutes a possible solution for minimally invasive cell and scaffold simultaneous delivery, but the reduced pore dimension and pore interconnection diameter of several commercially available MCs limits de facto cell ingrowth, and ultimately their suitability for in vivo cell delivery. In this study we investigated the potential of a new macroporous MC based on a collagen I-based recombinant peptide (Cellnest (TM)) for C2C12 cells and human bone marrow-derived mesenchymal stromal cells (hBMSCs) expansion and we analyzed the influence of dehydrothermal (DHT), hexamethylene diisocyanate (HMDIC), and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) cross-linking strategies on cell vitality, proliferation, and hBMSCs differentiation. We established a double emulsification protocol for the manufacturing of MCs characterized by external pores of 20-40 mu m diameter, 73% porosity, and 20-3 mu m pore interconnection diameter supporting cell ingrowth and proliferation into the MC. MCs cross-linked with DHT and HMDIC supported higher cell proliferation comparing to a commercially available equivalent over the course of 7 days and resulted in higher cell yield by day 28. Moreover, while hBMSCs expansion on Cellnest-MCs did not lead to a significant upregulation of the early markers of osteogenic differentiation Col1a1 and Runx2, their differentiation potential into the osteogenic lineage was preserved when cultured in differentiation medium, as confirmed by mineralized ECM deposition. We believe that Cellnest-MCs will help in reaching clinically relevant cell quantities and ultimately help in accelerating the translation of cell-based therapies for bone tissue engineering in the clinical practice.
机译：基于细胞外基质（ECM） - 丝合成材料的支架配方的发展构成了骨组织工程方法中基于细胞的疗法的进展的重要资源，其中通常需要细胞和支架植入。多孔微载体上的培养细胞（MCS）允许细胞膨胀在三维微观环境中，并构成可侵入性细胞和支架同时递送的可能溶液，但是几种商业上可获得的MCS的孔尺寸和孔隙互连直径的降低，最终适用于体内细胞递送。在该研究中，我们研究了基于C2C12细胞和人骨髓间充质基质细胞（HBMSCs）扩张的基于胶原I基重组肽（Cellnest（TM））的新大孔MC的潜力，并分析了脱水热量的影响（DHT），六亚甲基二异氰酸酯（HMDIC）和1-乙基-3-（3-二甲基氨基丙基）碳二亚胺（EDC）的交联策略对细胞活力，增殖和HBMSCs分化的交联策略。我们建立了一种双乳化方案，用于制造MCS，其外部孔径为20-40μm，孔隙率为73％和20-3μm孔互连直径，支撑电池发热和增殖进入MC。与DHT和HMDIC交联的MCS支持的更高细胞增殖与7天内的商业上可获得的相当，并导致28天的细胞产量增加。此外，在Cellnest-MC上的HBMSCs扩增不会导致显着的上调在骨质发生分化COL1A1和RUNX2的早期标记中，在分化介质中培养时，将它们的分化电位保持在成骨谱系中，如矿化ECM沉积的确认。我们认为Cellnest-MCS将有助于临床相关的细胞量，并最终有助于加速临床实践中骨组织工程的基于细胞的疗法的翻译。

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