Optimization of in vitro cell culture conditions suitable for the cholangiocarcinoma stem cell study
Year: 2025
Type: article
Abstract: Currently, cancer stem cells (CSCs) are extensively studied due to their roles in tumorigenesis, metastasis, and drug resistance.However, the low quantity, and its transient nature present a challenge for their study.Therefore, a culture system supporting high CSC quantity and flexible for high-throughput drug testing is needed.In this study, we used cholangiocarcinoma (CCA) cell lines with a CSC-specific live-cell biosensor system, which expresses green fluorescence protein (GFP) in CSCs, to study factors/conditions affecting CCA stem-like cell quantity in culture.We investigated culture media, seeding density, and compared 2-dimension (2-D) vs 3-dimensional (3-D) culture methods.The effects of anti-CCA drug dosages on CSC numbers were also studied.By comparing three media recipes, a defined serum-free CCA medium was selected for its high CSC yields.We found seeding density and time significantly influence CCA CSC percentages in both 2-D and 3-D settings.Notably, 3-D culture markedly increased CSC percentages compared to 2-D, supporting its application in CSC studies.In 3-D culture, a higher CSC index correlated with higher anti-CCA drug doses, suggesting loss of differentiation as a prominent characteristic under standard therapy.Thus, we report culture conditions influencing CCA CSC quantity, useful for CSC study and anti-CSC drug discovery in CCA.
Source: ScienceAsia
Institutions: Siriraj Hospital, Mahidol University, University of Minnesota
Language: English
Topic: Cancer Cells and Metastasis
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: diamond
Award: G5835890583
OpenAlex ID: W4412376394