Bio-fabrication of Liver Cancer Stem Cells and Testing of Gemcitabine

Aim: Cancer stem cells are cell populations that are essential in drug resistance and cancer metastasis. Some liver cancer cells exhibit the characteristics of cancer stem cells, and it is crucial to study the activities and interactions of drugs in these cells. Huh7 is a human liver cancer cell with stem cell biomarkers and is used with induced pluripotent stem cells to form various cancer organoids through encapsulation methods. Due to their ease of use without animal testing, bio-fabrication studies of cell-encapsulated models have gained importance in the pharmaceutical industry in recent years. This study aimed to biofabricate Huh7 human liver cancer stem cells by alginate encapsulation and test gemcitabine’s efficacy. Methods: Huh7 cells were encapsulated with alginate (0.8% w/v) and fibronectin, and their viability was evaluated with 3.2 µM gemcitabine on days 1, 3, 6, 9, and 12. Furthermore, gene expressions of stem cell markers CD90 and AFP were evaluated in encapsulated Huh7 cells by qPCR. In addition, IL-6 secretion in the cell medium was measured by ELISA for the tumor microenvironment. Results: Encapsulation of Huh7 cells was found to maintain their viability and stem cell properties for up to 12 days. In addition, alginate-encapsulated Huh7 cells were bio-fabricated to demonstrate long-term gemcitabine response. While the effect of the gemcitabine was evaluated in alginate-encapsulated Huh7 cells, CD90 and AFP mRNA levels were significantly reduced in the cells and IL-6 secretion was decreased in the tumor microenvironment. Conclusion: This study demonstrated that bio-fabrication of alginate-encapsulated Huh7 cells is a novel approach for long-term drug testing in liver cancer models. Bio-fabricated alginate-encapsulated cancer stem cells may be a cheaper and faster method for the testing of many drugs.