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D. Hokka



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    P3.01 - Poster Session with Presenters Present (ID 469)

    • Event: WCLC 2016
    • Type: Poster Presenters Present
    • Track: Biology/Pathology
    • Presentations: 1
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      P3.01-055 - In vitro Construction of Lung Cancer Organoids from Induced Lung Cancer Stem Like Cells (ID 4060)

      14:30 - 14:30  |  Author(s): D. Hokka

      • Abstract
      • Slides

      Background:
      Lung cancer stem cells are considered to be responsible for lung cancer progression. However, little is known about how they actually promote lung cancer progression and metastasis.

      Methods:
      We retrovirally introduced three defined factors (OCT3/4, SOX2, and KLF4) into lung cancer cell line, A549. We evaluated cancer stem cell properties in the A549 cells transduced with the three factors (OSK-A549) in terms of their chemo resistance, and sphere formation ability. We also assessed lung cancer organoid constructing ability by co-culturing with mesenchymal stem cells (MSC) and human umbilical vein epithelial cells (HUVEC).

      Results:
      OSK-A549 cells formed dome-shaped colonies in 10 to 15 days after transfection. These colonies were picked up for further expansion in DMEM/10%FBS medium, and we named these cells OSK-A549-Colony cells. Induced OSK-A549-colony cells were more resistant to cisplatin than parental A549 cells. Cell cycle analysis revealed that the rate of G0/G1 cells was significantly increased in OSK-A549-colony cells. Sphere forming ability was enhanced in OSK-A549-colony cells. These results suggested that OSK-A549-colony cells acquired the properties of lung cancer stem like cells. Co-culture with MSC and HUVEC showed that A549 and OSK-A549-colony cells could form large spheres equally, however, HE staining of spheres revealed that OSK-A549-colony cells could form much denser spheres than those of parental cells (Figure). As the morphology was similar to real lung cancer tissue, we named this spheres “lung cancer organoids”.Figure 1



      Conclusion:
      By introducing defined factors, A549 cells acquired lung cancer stem cell like properties, and these cells could form lung cancer organoids by co-culturing with MSC and HUVEC. Analysis of these organoids might enable us to elucidate the molecular mechanism of lung cancer progression and metastasis.

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