Further characterization of human fetal osteoblastic hFOB 1.19 and hFOB/ERα cells:: bone formation in vivo and karyotype analysis using multicolor fluorescent in situ hybridization
Further characterization of human fetal osteoblastic hFOB 1.19 and hFOB/ERα cells:: bone formation in vivo and karyotype analysis using multicolor fluorescent in situ hybridization
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DOI：
10.1002/jcb.10259
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发表时间：
2002-01-01
影响因子：
4
通讯作者：
Spelsberg, TC
中科院分区：
文献类型：
--
作者：
Subramaniam, M;Jalal, SM;Spelsberg, TC
我们曾利用稳定转染的温度敏感的SV40 t抗原，生成了永生化的人胎儿成骨细胞系（hFOB） (Harris etal . [1995a]。矿工。> 10:178 - 1860)。为了表征这些细胞的表型/基因型属性，以获得良好的细胞模型系统，我们通过多色荧光原位杂交（M-FISH）进行了核型分析，分析了它们在体内形成骨而不发生细胞转化的能力，最后分析了它们在体外形成细胞外基质的能力。hFOB细胞的核型分析显示1-2条染色体的结构或数量异常。相反，MG63人骨肉瘤细胞表现出多种，通常是复杂的，数量和结构异常。在Matrigel存在的情况下皮下注射hFOB细胞至裸鼠2-3周后骨形成。培养hFOB细胞沉积的细胞外基质的电镜分析显示，平行排列的轻带状原纤维是I型和III型纤维胶原的典型特征。这些结果表明，hFOB细胞系具有最小的染色体异常，表现出分化成骨细胞的基质合成特性，是永生化但非转化的细胞系。因此，这些hFOB细胞似乎是体外研究成骨细胞生物学的一个很好的模型系统。j .细胞。生物化学学报，21(3):397 - 398。(C) 2002 Wiley-Liss, Inc。
We have previously generated an immortalized human fetal osteoblastic cell line (hFOB) using stably transfected temperature sensitive SV40 T-antigen (Harris et al. [ 1995a] J. Bone. Miner. Res. 10:178-1860). To characterize these cells for phenotypic/genotypic attributes desired for a good cell model system, we performed karyotype analysis by multicolor fluorescent in situ hybridization (M-FISH), their ability to form bone in vivo without developing cell transformation, and finally their ability to form extracellular matrix formation in vitro. The karyotype analysis of hFOB cells revealed structural or numeric anomalies involving 1-2 chromosomes. In contrast, the human osteosarcoma MG63 cells displayed multiple, and often complex, numeric, and structural abnormalities. Subcutaneous injection of hFOB cells in the presence of Matrigel into nude mice resulted in bone formation after 2-3 weeks. Electron microscopic analysis of the extracellular matrix deposited by hFOB cells in culture revealed a parallel array of lightly banded fibrils typical of the fibrillar collagens such as type I and III. These results demonstrate that the hFOB cell line has minimal chromosome abnormalities, exhibit the matrix synthetic properties of differentiated osteoblasts, and are immortalized but non-transformed cell line. These hFOB cells thus appear to be an excellent model system for the study of osteoblast biology in vitro. J. Cell. Biochem. 87: 9-15, 2002. (C) 2002 Wiley-Liss, Inc.