Optimization of a chondrogenic medium through the use of factorial design of experiments.
Optimization of a chondrogenic medium through the use of factorial design of experiments.
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DOI：
10.1089/biores.2012.0277
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发表时间：
2012-12
影响因子：
--
通讯作者：
Lindahl A
中科院分区：
文献类型：
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作者：
Enochson L;Brittberg M;Lindahl A
用于体外软骨研究的标准培养系统基于三维微团培养中的细胞和含有软骨形成关键生长因子转化生长因子（TGF）-β1的限定培养基。本研究旨在优化软骨细胞微团培养的培养基。人软骨细胞在不同的培养基配方中培养，这些培养基配方采用析因实验设计（DoE）方法设计，并基于用于再分化的标准培养基。通过响应面分析法，对软骨细胞再分化的影响因素进行了优化。TGF-β1、地塞米松和葡萄糖是促进软骨细胞分化的重要因素。与标准培养基相比，TGF-β1增加了30%，地塞米松减少了50%，葡萄糖增加了22%。在与标准培养基的比较研究中验证了优化培养基的效价。优化的培养基导致微团培养物中对关节软骨细胞表型重要的基因表达增加，并且培养物中糖胺聚糖/DNA含量增加。采用高效DoE法对标准培养基进行优化，确定了一种对关节软骨细胞再分化效果更好的新培养基。
The standard culture system for in vitro cartilage research is based on cells in a three-dimensional micromass culture and a defined medium containing the chondrogenic key growth factor, transforming growth factor (TGF)-β1. The aim of this study was to optimize the medium for chondrocyte micromass culture. Human chondrocytes were cultured in different media formulations, designed with a factorial design of experiments (DoE) approach and based on the standard medium for redifferentiation. The significant factors for the redifferentiation of the chondrocytes were determined and optimized in a two-step process through the use of response surface methodology. TGF-β1, dexamethasone, and glucose were significant factors for differentiating the chondrocytes. Compared to the standard medium, TGF-β1 was increased 30%, dexamethasone reduced 50%, and glucose increased 22%. The potency of the optimized medium was validated in a comparative study against the standard medium. The optimized medium resulted in micromass cultures with increased expression of genes important for the articular chondrocyte phenotype and in cultures with increased glycosaminoglycan/DNA content. Optimizing the standard medium with the efficient DoE method, a new medium that gave better redifferentiation for articular chondrocytes was determined.