Goldring Mary B
Division of Rheumatology, Beth Israel Deaconess Medical Center, New England Baptist Bone and Joint Institute, Harvard Institutes of Medicine, Boston, MA, USA.
Methods Mol Med. 2004;100:37-52. doi: 10.1385/1-59259-810-2:037.
Immortalization of chondrocytes increases life span and proliferative capacity but does not necessarily stabilize the differentiated phenotype. Expansion of chondrocyte cell lines in continuous monolayer culture may result in the loss of phenotype, particularly if high cell density is not maintained. This chapter describes strategies for maintaining or restoring differentiated phenotype in established chondrocyte cell lines involving culture in serum-free defined culture medium, in suspension over agarose or polyHEMA, or within alginate or collagen scaffolds. Chondrocyte cell lines have been used successfully to develop reproducible models for studying the regulation of gene expression in experiments requiring large numbers of cells. Thus, approaches for studying transcriptional regulation by transfection of promoter-driven reporter genes and cotransfection of expression vectors for wild-type or mutant proteins are also described.
软骨细胞永生化可延长其寿命并提高增殖能力，但不一定能稳定其分化表型。在连续单层培养中扩增软骨细胞系可能导致表型丧失，尤其是在未维持高细胞密度的情况下。本章介绍了在已建立的软骨细胞系中维持或恢复分化表型的策略，包括在无血清限定培养基中培养、在琼脂糖或聚甲基丙烯酸羟乙酯上悬浮培养，或在藻酸盐或胶原蛋白支架内培养。软骨细胞系已成功用于建立可重复的模型，以在需要大量细胞的实验中研究基因表达的调控。因此，还介绍了通过转染启动子驱动的报告基因以及共转染野生型或突变蛋白表达载体来研究转录调控的方法。
