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源自人软骨肉瘤的具有软骨细胞表型的新细胞系。

New cell lines with chondrocytic phenotypes from human chondrosarcoma.

作者信息

Kudawara Ikuo, Araki Nobuhito, Myoui Akira, Kato Yoichi, Uchida Atsumasa, Yoshikawa Hideki

机构信息

Department of Orthopaedic Surgery, Osaka National Hospital, Osaka 2-1-14 Hoenzaka, Chuo-ku, 540-0006 Osaka, Japan.

出版信息

Virchows Arch. 2004 Jun;444(6):577-86. doi: 10.1007/s00428-004-1020-6. Epub 2004 Apr 29.

Abstract

In the present study, we investigated chondrocytic characterization for newly established human chondrosarcoma cell lines. A chondrosarcoma cell line, HCS-TG, was established by the implantation of grade-2 human chondrosarcoma into athymic mice. Cloning of HCS-TG cells from passage 17 was performed. After cell cloning, two clonal-cell lines (HCS-TG C3 and E2) with good proliferative activities were obtained. These cell lines in monolayer culture retained a polygonal morphology. Their doubling times were 68 h and 45 h, respectively. mRNA expressions of type-I, -II, -X, and -XI collagens and aggrecan core protein were detected on reverse-transcription polymerase chain reaction. Protein expression of type-II collagen was confirmed in each cell line using Western blotting. However, there was no expression of type-I collagen. Moreover, gelatin zymography revealed that both cell lines produced extracellular matrices with matrix metalloproteinases 2 and 9. The parental HCS-TG cells had tumorigenicity in athymic mice; however, C3 and E2 were not tumorigenic. New clonal-cell lines HCS-TG C3 and E2 derived from human chondrosarcoma are morphologically chondrocytic in serial monolayer cultures and express chondrocytic phenotypes.

摘要

在本研究中,我们对新建立的人软骨肉瘤细胞系进行了软骨细胞特性研究。通过将2级人软骨肉瘤植入无胸腺小鼠体内,建立了软骨肉瘤细胞系HCS-TG。对第17代的HCS-TG细胞进行克隆。细胞克隆后,获得了两个具有良好增殖活性的克隆细胞系(HCS-TG C3和E2)。这些单层培养的细胞系保持多边形形态。它们的倍增时间分别为68小时和45小时。通过逆转录聚合酶链反应检测了I型、II型、X型和XI型胶原蛋白以及聚集蛋白聚糖核心蛋白的mRNA表达。使用蛋白质印迹法在每个细胞系中证实了II型胶原蛋白的蛋白表达。然而,未检测到I型胶原蛋白的表达。此外,明胶酶谱分析表明,这两个细胞系都产生含有基质金属蛋白酶2和9的细胞外基质。亲代HCS-TG细胞在无胸腺小鼠中具有致瘤性;然而,C3和E2没有致瘤性。源自人软骨肉瘤的新克隆细胞系HCS-TG C3和E2在连续单层培养中形态上呈软骨细胞样,并表达软骨细胞表型。

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