P3.01-055 In vitro Construction of Lung Cancer Organoids from Induced Lung Cancer Stem Like Cells: Topic: Stem Cells in Lung Cancer

Abstract

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. 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). 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”. 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.