Alegre-Aguarón Elena, Sampat Sonal R, Xiong Jennifer C, Colligan Ryan M, Bulinski J Chloë, Cook James L, Ateshian Gerard A, Brown Lewis M, Hung Clark T
Department of Biomedical Engineering, Columbia University, New York, New York, United States of America.
Quantitative Proteomics Center, Columbia University, New York, New York, United States of America.
PLoS One. 2014 Feb 7;9(2):e88053. doi: 10.1371/journal.pone.0088053. eCollection 2014.
To make progress in cartilage repair it is essential to optimize protocols for two-dimensional cell expansion. Chondrocytes and SDSCs are promising cell sources for cartilage repair. We previously observed that priming with a specific growth factor cocktail (1 ng/mL transforming growth factor-β1, 5 ng/mL basic fibroblast growth factor, and 10 ng/mL platelet-derived growth factor-BB) in two-dimensional culture, led to significant improvement in mechanical and biochemical properties of synovium-derived stem cell (SDSC)-seeded constructs. The current study assessed the effect of growth factor priming on the proteome of canine chondrocytes and SDSCs. In particular, growth factor priming modulated the proteins associated with the extracellular matrix in two-dimensional cultures of chondrocytes and SDSCs, inducing a partial dedifferentiation of chondrocytes (most proteins associated with cartilage were down-regulated in primed chondrocytes) and a partial differentiation of SDSCs (some collagen-related proteins were up-regulated in primed SDSCs). However, when chondrocytes and SDSCs were grown in pellet culture, growth factor-primed cells maintained their chondrogenic potential with respect to glycosaminoglycan and collagen production. In conclusion, the strength of the label-free proteomics technique is that it allows for the determination of changes in components of the extracellular matrix proteome in chondrocytes and SDSCs in response to growth factor priming, which could help in future tissue engineering strategies.
为了在软骨修复方面取得进展，优化二维细胞扩增方案至关重要。软骨细胞和滑膜衍生干细胞（SDSCs）是软骨修复中很有前景的细胞来源。我们之前观察到，在二维培养中用特定生长因子鸡尾酒（1 ng/mL转化生长因子-β1、5 ng/mL碱性成纤维细胞生长因子和10 ng/mL血小板衍生生长因子-BB）进行预处理，可显著改善滑膜衍生干细胞（SDSC）接种构建体的力学和生化特性。本研究评估了生长因子预处理对犬软骨细胞和SDSCs蛋白质组的影响。特别是，生长因子预处理在软骨细胞和SDSCs的二维培养中调节了与细胞外基质相关的蛋白质，诱导软骨细胞部分去分化（在预处理的软骨细胞中，大多数与软骨相关的蛋白质下调）和SDSCs部分分化（在预处理的SDSCs中，一些与胶原蛋白相关的蛋白质上调）。然而，当软骨细胞和SDSCs在微团培养中生长时，生长因子预处理的细胞在糖胺聚糖和胶原蛋白产生方面保持其软骨生成潜力。总之，无标记蛋白质组学技术的优势在于它能够确定软骨细胞和SDSCs中细胞外基质蛋白质组成分响应生长因子预处理的变化，这可能有助于未来的组织工程策略。
