Wu Xiaoyuan, Zhu Lei, Xia Lifeng, Gui Lin, Cong Wei
School of Stomatology, Dalian Medical University, Dalian, Liaoning, China.
Department of Periodontics, Affiliated Stomatological Hospital of Suzhou Vocational Health College, Suzhou, Jiangsu, China.
Regen Ther. 2025 Mar 13;29:108-116. doi: 10.1016/j.reth.2025.02.003. eCollection 2025 Jun.
FAM20B is a newly identified kinase involved in proteoglycan biosynthesis. Inactivation of in joint cartilage leads to chondrosarcoma. Generation of chondrocytes cell line carrying floxed allele can offer a valuable tool for the study of the role of as well as the molecular events in chondrocyte biology and cartilage diseases. The limitations in the primary culture of chondrocytes necessitate the development of chondrocyte cell line. In this study, we established and characterized the immortalized mouse floxed chondrocyte cell line. The primary mouse floxed chondrocytes were isolated from the articular cartilage of knee joints and immortalized using lentivirus containing Simian Virus 40 T-antigen (SV40 T-Ag). The immortalized cell line was verified by genomic integration of SV40 T-Ag and proliferated at a high rate relative to their primary counterparts. The immortalized chondrocyte cell line not only retained the typical ultrastructural morphology and important phenotypic characteristics of articular cartilage, but also possessed strong differentiation potential upon three-dimensional pellet culture. Thus, we, for the first time, describe the development of immortalized mouse floxed chondrocytes and present an alternative for the limited number of articular chondrocyte cell lines for the study of cartilage biology.
FAM20B是一种新发现的参与蛋白聚糖生物合成的激酶。关节软骨中FAM20B的失活会导致软骨肉瘤。构建携带FAM20B基因floxed等位基因的软骨细胞系可为研究FAM20B的作用以及软骨细胞生物学和软骨疾病中的分子事件提供有价值的工具。软骨细胞原代培养的局限性使得软骨细胞系的建立成为必要。在本研究中，我们建立并鉴定了永生化的小鼠FAM20B基因floxed软骨细胞系。原代小鼠FAM20B基因floxed软骨细胞从膝关节的关节软骨中分离出来，并使用含有猿猴病毒40 T抗原（SV40 T-Ag）的慢病毒进行永生化处理。通过SV40 T-Ag的基因组整合验证了永生化细胞系，并且相对于其原代细胞以高增殖速率增殖。永生化软骨细胞系不仅保留了关节软骨典型的超微结构形态和重要的表型特征，而且在三维微球培养时具有很强的分化潜能。因此，我们首次描述了永生化小鼠FAM20B基因floxed软骨细胞的构建，并为软骨生物学研究中数量有限的关节软骨细胞系提供了一种替代选择。
