逆转录病毒介导的癌基因转导嗅上皮细胞的细胞系

Calof Anne L., Guevara Jose L.
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引用次数: 9

摘要

我们描述了从E15小鼠胚胎中纯化的嗅上皮(OE)原代培养物中获得的六种永生细胞系的分离和表征。使用含有人c-myc cDNA以及显性选择性抗生素抗性基因(neo)的复制缺陷鼠逆转录病毒使培养的细胞永生化。扩增并分析在抗生素选择中存活下来并显示出带过程形态的细胞,以确定嗅觉受体神经元、支持细胞和嗅觉鞘细胞特征的分子标记物的表达。有趣的是,所有六种细胞系都表达了嗅神经鞘细胞或施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标志物。我们的研究结果表明,虽然使用逆转录病毒介导的癌基因转移可以产生永生化OE细胞系,但可以永生化的细胞类型可能存在局限性。此外,我们证明了永生化OE细胞系在启动子陷阱实验中鉴定发育调控基因的潜在有用性。
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Cell Lines Derived from Retrovirus-Mediated Oncogene Transduction into Olfactory Epithelium Cultures

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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