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首页> 外文期刊>Academic Journal of Xi'an Jiaotong University >Construction and characterization of hGM-CSF-expressing K-562 cell line
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Construction and characterization of hGM-CSF-expressing K-562 cell line

机译：表达hGM-CSF的K-562细胞系的构建与鉴定

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Objective The whole process of vaccine preparation is time-consuming and technically challenging. Here the hGM-CSF-engineered K-562 cell line was constructed to simplify tumor vaccine preparation process. Methods The eukaryocyte expressing plasmid pcDNA3. 1/GM-CSF was first constructed and its accuracy was verified through sequencing. The pcDNA3. 1/GM-CSF was transfected into COS-7 cells to verify GM-CSF expression and cytokine activity using TF-1 cell line. Then the plasmid was transfected into K-562 cell line using liposome method, and was selected under G-418 and sub-cloned by limiting dilution. GM-CSF product from the monoclone GM-CSF-K-562 cell lines was quantified using ELISA method. Results We successfully constructed the hGM-CSF eukaryocyte expressing plasmid and hGM-CSF expressing K-562 cell line. Conclusion The construction of K-562/GM-CSF line will simplify the preparation of tumor vaccine, thus facilitating the application of tumor vaccination therapy in clinical application.
机译：目的疫苗制备的整个过程既耗时又具有技术挑战。在这里，构建了由hGM-CSF设计的K-562细胞系，以简化肿瘤疫苗的制备过程。方法真核细胞表达质粒pcDNA3。首先构建1 / GM-CSF，并通过测序验证其准确性。 pcDNA3。将1 / GM-CSF转染到COS-7细胞中，以使用TF-1细胞系验证GM-CSF表达和细胞因子活性。然后用脂质体方法将质粒转染到K-562细胞系中，在G-418下选择并通过有限稀释亚克隆。使用ELISA方法对来自单克隆GM-CSF-K-562细胞系的GM-CSF产物进行定量。结果成功构建了表达hGM-CSF真核细胞的质粒和表达hGM-CSF的K-562细胞。结论K-562 / GM-CSF细胞系的构建将简化肿瘤疫苗的制备，有利于肿瘤疫苗治疗在临床中的应用。

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