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Liver Cancer Stem Cell Induction from Induced Pluripotent Stem Cells
( Said M Afify ),( Ghmkin Hassan ),( Hend M Nawara ),( Hager M Mansour ),( Amira Osman ),( Sadia Monzur ),( Hagar Ali Abu Quora ),( Maram H Zahra ),( Akimasa Seno ),( Yoshiaki Iwasaki ),( Masaharu Sen 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1
Aims: Liver cancer stem cells represent a small fraction of cells in liver cancer tissues so that studying these cells is very hard. Generation of liver cancer stem cells considered as one of the most important issue in cancer biology research. For this reason, we tried to generate liver cancer stem cells from induced pluripotent stem (iPSCs). Methods: First of all, CM was collected from confluent culture of Huh7 cells. Then, mouse iPSCs cells without MEF feeder cells were cultured in the presence of 50% CM for 4 weeks. The medium was changed every day with fresh medium containing 50% of CM. Mouse iPSCs cultured is the complete medium with LIF were used as a control. The survived cells (5x105 cells) were suspended in HBSS and injected into the liver of BALB/ c nude mice. After 25 days malignant tumor was formed in the liver while benign teratoma was formed by the injection of iPSCs. Tumors were then excised and partly fixed in 10% neutral formalin buffer solution for HE staining and immunohistochemical analysis. The rest of tumors were subjected to rt-qPCR anaylsis and primary culture. Results: Immunohistochemical analysis with liver cancer associated markers and cancer stem cell marker showed that malignant liver tumor was developed. These results indicate that the primary cells from the malignant tumor are rich in CSCs. Conclusions: This model will be very important and useful to assess the significant molecular mechanisms necessary to maintain liver cancer stem cells, which will help in defat liver cancer.