Department of Orthopaedic Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
J Orthop Res. 2012 Oct;30(10):1666-73. doi: 10.1002/jor.22113. Epub 2012 Apr 13.
Chordoma is a rare primary malignant bone tumor and there exist only a few established human chordoma cell lines. The scarcity of robust chordoma cell lines has limited the ability to study this tumor. In this report, we describe the establishment of a novel chordoma cell line and characterize its in vitro and in vivo behaviors. The tumor tissue was isolated from a patient with recurrent chordoma of the sacrum. After 6 months in culture, the chordoma cell line, referred here as CH22, was established. Microscopic analysis of two-dimensional culture confirmed that the CH22 cells exhibited a typical vacuolated cytoplasm similar to the well-established chordoma cell line U-CH1. Electron microscopy showed cohesive cells with numerous surface filopodia, pockets of glycogen and aggregates of intermediate tonofilaments in cytoplasm. Three-dimensional culture revealed that the CH22 cells could grow and form clusters by day 8. The MTT assays demonstrated that, compared with sensitive osteosarcoma cell lines, CH22 cells were relatively resistant to conventional chemotherapeutic drugs. Western blotting and immunofluorescence analysis confirmed that the CH22 cells expressed brachyury, vimentin, and cytokeratin. Finally, histological analysis of CH22 xenograft tumor tissues demonstrated the appearance of physaliphorous cells and positive staining of brachyury, cytokeratin, and S100. By CT and MRI, imaging xenografts showed the typical appearances seen in human chordomas. These findings suggest that the established novel human chordoma cell line CH22 and its tumorigenecity in SCID nude mice may serve as an important model for studying chordoma cell biology and the development of new therapeutic modalities.
软骨肉瘤是一种罕见的原发性恶性骨肿瘤，目前仅有少数几种已建立的人类软骨肉瘤细胞系。由于缺乏稳健的软骨肉瘤细胞系，限制了对这种肿瘤的研究能力。在本报告中，我们描述了一种新型软骨肉瘤细胞系的建立，并对其体外和体内行为进行了特征描述。肿瘤组织取自一位复发性骶骨软骨肉瘤患者。经过 6 个月的培养，建立了软骨肉瘤细胞系，称为 CH22。二维培养的显微镜分析证实，CH22 细胞表现出典型的空泡状细胞质，类似于已建立的软骨肉瘤细胞系 U-CH1。电子显微镜显示，细胞间有许多表面丝状伪足，细胞质中有糖原小囊和中间张力丝聚集。三维培养显示，CH22 细胞在第 8 天可以生长并形成细胞簇。MTT 检测表明，与敏感的骨肉瘤细胞系相比，CH22 细胞对常规化疗药物相对耐药。Western blot 和免疫荧光分析证实，CH22 细胞表达 brachyury、vimentin 和 cytokeratin。最后，CH22 异种移植肿瘤组织的组织学分析显示出 physaliphorous 细胞的出现，以及 brachyury、cytokeratin 和 S100 的阳性染色。通过 CT 和 MRI，成像异种移植显示出人类软骨肉瘤中常见的典型外观。这些发现表明，建立的新型人类软骨肉瘤细胞系 CH22 及其在 SCID 裸鼠中的致瘤性可能成为研究软骨肉瘤细胞生物学和开发新治疗方法的重要模型。
