逆转录病毒介导癌基因转导至嗅上皮培养的细胞系

A. Calof, J. L. Guevara
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引用次数: 9

摘要

摘要:我们描述了从E15小鼠胚胎中纯化的嗅上皮(OE)原代培养的6株不朽细胞系的分离和特性。使用含有人c-myc cDNA的复制缺陷小鼠逆转录病毒,以及显性选择性抗生素抗性基因(neo),使培养的细胞永生化。对在抗生素选择中存活下来的细胞进行扩增,分析其嗅觉受体神经元、支撑细胞和嗅鞘细胞特征分子标记的表达。有趣的是,所有六种细胞系都表达了嗅神经鞘细胞或雪旺细胞的形态学和免疫学特性,但不表达嗅觉受体神经元的特征标记。我们的研究结果表明,虽然利用逆转录病毒介导的癌基因转移有可能产生永生化的OE细胞系,但可永生化的细胞类型可能存在限制。此外,我们证明了永生OE细胞系在启动子陷阱实验中鉴定发育调节基因的潜在用途。
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Cell Lines Derived from Retrovirus-Mediated Oncogene Transduction into Olfactory Epithelium Cultures
Abstract We describe the isolation and characterization of six immortal cell lines derived from primary cultures of olfactory epithelium (OE) purified from E15 mouse embryos. Cultured cells were immortalized using a replication-defective murine retrovirus containing the cDNA for human c-myc, in addition to a dominant selectable antibiotic resistance gene (neo). Cells that survived antibiotic selection and displayed process-bearing morphologies were expanded and analyzed for expression of molecular markers characteristic of olfactory receptor neurons, sustentacular cells, and olfactory ensheathing cells. Interestingly, all six cell lines expressed morphological and immunological properties of the ensheathing, or Schwann, cells of the olfactory nerve, but did not express markers characteristic of olfactory receptor neurons. Our results suggest that, while it is possible to generate immortalized OE cell lines using retrovirus-mediated oncogene transfer, there may be limitations to the types of cells that can be immortalized. In addition, we demonstrate the potential usefulness of immortalized OE cell lines for promoter-trap experiments to identify developmentally regulated genes.
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