Modulation of chondrogenic differentiation of human mesenchymal stem cells in jellyfish collagen scaffolds by cell density and culture medium
Modulation of chondrogenic differentiation of human mesenchymal stem cells in jellyfish collagen scaffolds by cell density and culture medium
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DOI：
10.1002/term.2065
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发表时间：
2017-06-01
影响因子：
3.3
通讯作者：
Bernhardt, A.
中科院分区：
文献类型：
--
作者：
Pustlauk, W.;Paul, B.;Bernhardt, A.
用于再生创伤软骨的组织工程方法的研究越来越关注多能人间充质干细胞（hMSC）作为自体软骨细胞的替代品。本研究应用由水母 Rhopilema esculentum 胶原蛋白制成的多孔支架进行 hMSC 的体外软骨分化。这些支架中的培养条件与高密度颗粒培养中的条件不同，因此有必要重新检查这些数据。我们系统地研究了接种密度、不同葡萄糖含量的基础培养基[杜尔伯科改良伊格尔培养基（DMEM）、最低必需培养基（-MEM）]以及添加胎牛血清（FCS）或牛血清白蛋白（BSA）对hMSCs软骨分化的影响。对软骨形成分化和肥大发育的选定标记物进行基因表达分析。此外，通过定量硫酸化糖胺聚糖和 II 型胶原蛋白含量来分析软骨细胞外基质 (ECM) 的产生。在高糖 DMEM 和 0.125% BSA 中培养后，以 2.4 x 10(6) 细胞/cm(3) 接种的支架中观察到软骨形成标记物的最强上调以及最高的 ECM 沉积。与高密度颗粒培养物相比，较低的接种密度足以诱导胶原支架中 hMSC 的体外软骨形成，这减少了支架接种所需的细胞量，从而减少了单层扩增周期。此外，检查FCS和-MEM对软骨形成的MSC分化的影响是开发用于骨软骨组织再生的双相支架中同时成骨和软骨形成的骨软骨介质的重要先决条件。版权所有 (c) 2015 约翰·威利父子有限公司
Studies on tissue-engineering approaches for the regeneration of traumatized cartilage focus increasingly on multipotent human mesenchymal stem cells (hMSCs) as an alternative to autologous chondrocytes. The present study applied porous scaffolds made of collagen from the jellyfish Rhopilema esculentum for the in vitro chondrogenic differentiation of hMSCs. Culture conditions in those scaffolds differ from conditions in high-density pellet cultures, making a re-examination of these data necessary. We systematically investigated the influence of seeding density, basic culture media [Dulbecco's modified Eagle's medium (DMEM), -minimum essential medium (-MEM)] with varying glucose content and supplementation with fetal calf serum (FCS) or bovine serum albumin (BSA) on the chondrogenic differentiation of hMSCs. Gene expression analyses of selected markers for chondrogenic differentiation and hypertrophic development were conducted. Furthermore, the production of cartilage extracellular matrix (ECM) was analysed by quantification of sulphated glycosaminoglycan and collagen type II contents. The strongest upregulation of chondrogenic markers, along with the highest ECM deposition was observed in scaffolds seeded with 2.4 x 10(6) cells/cm(3) after cultivation in high-glucose DMEM and 0.125% BSA. Lower seeding densities compared to high-density pellet cultures were sufficient to induce in vitro chondrogenic differentiation of hMSCs in collagen scaffolds, which reduces the amount of cells required for the seeding of scaffolds and thus the monolayer expansion period. Furthermore, examination of the impact of FCS and -MEM on chondrogenic MSC differentiation is an important prerequisite for the development of an osteochondral medium for simultaneous osteogenic and chondrogenic differentiation in biphasic scaffolds for osteochondral tissue regeneration. Copyright (c) 2015 John Wiley & Sons, Ltd.