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Establishing a lung cancer stem cell culture using autologous intratumoral fibroblasts as feeder cells

Authors

  • Yu Xu,

    1. The Third Department of Oncology, PLA Cancer Research Institute of the Second Affiliated Hospital, The Third Military Medical University, ChongQing 400037, Peoples Republic of China
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  • Yi-De Hu,

    Corresponding author
    1. The Third Department of Oncology, PLA Cancer Research Institute of the Second Affiliated Hospital, The Third Military Medical University, ChongQing 400037, Peoples Republic of China
    2. The Third Department of Oncology, XinQiao Hospital, ShaPingBa District, Chongqing 400037, Peoples Republic of China
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  • Jing Zhou,

    1. The Third Department of Oncology, PLA Cancer Research Institute of the Second Affiliated Hospital, The Third Military Medical University, ChongQing 400037, Peoples Republic of China
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  • Ming-Hui Zhang

    1. The Third Department of Oncology, PLA Cancer Research Institute of the Second Affiliated Hospital, The Third Military Medical University, ChongQing 400037, Peoples Republic of China
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To whom correspondence should be addressed (email huyide_mit@yahoo.com.cn).

Abstract

Human LCSCs (lung cancer stem cells) were first isolated from lung cancer patients and cultured using serum-free culture methods. To recreate the intratumoural microenvironment to sustain LCSC growth, autologous intratumoral fibroblasts were used as feeder cells. In this study, we investigated the growth and maintenance of pluripotency in prolonged LCSCs culture on autologous intratumoural fibroblasts. LCSCs isolated from three clinical samples all showed vigorous growth on feeder cells for 16 weeks of continuous cultures with a doubling time of 41–47 h. The cells continued expressing stem cell marker CD133 and remained undifferentiated. Pluripotency was demonstrated by tumour formation in immunodeficient mice. In a feeder-free culture system, growth of LCSCs spheres was retarded and would cease when the diameter reached 100 μm if immediate passage was not performed. Moreover, spontaneous differentiation was more frequently seen in a serum-free culture system. In conclusion, we have successfully established a culture system using autologous intratumoural fibroblast cells as feeder cells for prolonged culture of undifferentiated LCSCs in vitro.

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