Tissue Regeneration Materials Unit, International Center for Materials Nanoarchitectonics, National Institute for Materials Science, Tsukuba, Ibaraki 305-0044, Japan.
J Biomed Mater Res A. 2012 Mar;100(3):694-702. doi: 10.1002/jbm.a.34003. Epub 2011 Dec 30.
The loss of cartilaginous phenotype during in vitro expansion culture of chondrocytes is a major barrier for the application of cartilage tissue engineering. The use of matrices mimicking the in vivo extracellular matrix (ECM) microenvironment is anticipated to be an efficient method to suppress chondrocyte phenotype loss. In this study, we developed several types of ECM derived from serially passaged chondrocytes for use as cell-culture substrata and compared their effects on chondrocyte functions. Primary bovine chondrocytes and serially passaged chondrocytes (at passages 2 and 6) were cultured on tissue-culture polystyrene. After culture, the cellular components were selectively removed from the ECM deposited by the cells. The remaining ECM proteins were used as cell-culture substrata. The composition of the deposited ECM depended on the culture stage of the serially passaged chondrocytes used for the ECM production. The deposited ECM supported the adhesion and proliferation of chondrocytes. The effects of the ECM on the chondrocyte dedifferentiation during in vitro passage culture differed dramatically depending on the phenotype of the chondrocytes used to produce the ECM. The primary chondrocyte-derived ECM delayed the chondrocyte dedifferentiation during in vitro passage culture and is a good candidate for chondrocyte subculture for tissue engineering.
软骨细胞在体外扩增培养过程中软骨表型的丧失是软骨组织工程应用的主要障碍。预计使用模拟体内细胞外基质（ECM）微环境的基质是抑制软骨细胞表型丧失的有效方法。在这项研究中，我们开发了几种源自传代软骨细胞的 ECM 用于作为细胞培养底物，并比较了它们对软骨细胞功能的影响。原代牛软骨细胞和传代软骨细胞（第 2 代和第 6 代）在组织培养聚苯乙烯上培养。培养后，从细胞分泌的 ECM 中选择性地去除细胞成分。剩余的 ECM 蛋白被用作细胞培养底物。沉积的 ECM 的组成取决于用于产生 ECM 的传代软骨细胞的培养阶段。沉积的 ECM 支持软骨细胞的黏附和增殖。ECM 对体外传代培养过程中软骨细胞去分化的影响因用于产生 ECM 的软骨细胞的表型而异。原代软骨细胞衍生的 ECM 延迟了体外传代培养过程中软骨细胞的去分化，是组织工程中软骨细胞亚培养的良好候选物。
