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Updated: Jul 4, 2025
Microfluidic Co-culture of Renal Healthy and Tumor Epithelium to Model Kidney Cancer Progression
Published on: January 31, 2025
Selective expansion of renal cancer stem cells using microfluidic single-cell culture arrays for anticancer drug
Xiaogang Wang1, Tao He1, Zihe Chen2
1Department of Urology, The Second Affiliated Hospital, Dalian Medical University, 467, Zhongshan Road, Shahekou District, Dalian 116021, Liaoning, China. lixiancheng@dmu.edu.cn.
Abstract:
The suboptimal prognosis associated with drug therapy for renal cancer can be attributed to the presence of stem-cell-like renal cancer cells. However, the limited number of these cells prevents conventional drug screening assays from effectively assessing the response of renal cancer stem cells to anti-cancer agents. To address this issue, the present study employed microfluidic single-cell culture arrays to expand renal cancer stem cells by exploiting the anti-apoptosis and self-renewal properties of tumor stem cells. A microfluidic chip with 18 000 hydrophilic microwells was designed and fabricated to establish the single-cell culture array. Over a 7 day culture, the large-scale single-cell culture yielded a limited quantity of single-cell-derived tumorspheres. The sphere formation rates for Caki-1, 786-O, and ACHN cells were determined to be 8.74 ± 0.53%, 12.02 ± 1.43%, and 4.98 ± 1.68%, respectively. The expanded cells exhibited stemness characteristics, as indicated by immunofluorescence, flow cytometry, serial passaging, and in vitro differentiation assays. Additionally, the comparative transcriptomic analysis showed significant differences in the gene expression patterns of the expanded cells compared to the differentiated renal cancer cells. The drug testing indicated that renal cancer stem cells exhibited reduced sensitivity towards the tyrosine kinase inhibitors sorafenib and sunitinib, compared to differentiated renal cancer cells. This reduced sensitivity can be attributed to the elevated expression levels of tyrosine kinase in renal cancer stem cells. This present study provides evidence that the utilization of microfluidic single-cell culture arrays for selective cell expansion can facilitate drug testing of renal cancer stem cells.
Insights
Microfluidic technology enables the expansion of rare renal cancer stem cells. This advancement allows for effective drug screening, revealing their reduced sensitivity to tyrosine kinase inhibitors.
Area of Science:
- Oncology
- Biotechnology
- Biomedical Engineering
Background:
- Renal cancer drug therapy is often suboptimal due to the presence of stem-cell-like cancer cells.
- Limited numbers of these cells hinder effective drug screening and response assessment.
- Tumor stem cells possess anti-apoptosis and self-renewal properties crucial for expansion.
Purpose of the Study:
- To develop a method for expanding renal cancer stem cells using microfluidic technology.
- To assess the drug sensitivity of expanded renal cancer stem cells.
- To investigate the gene expression differences between stem and differentiated renal cancer cells.
Main Methods:
- Fabrication of a microfluidic chip with 18,000 hydrophilic microwells for single-cell culture.
- Expansion of renal cancer stem cells over a 7-day culture period.
- Characterization of expanded cells using immunofluorescence, flow cytometry, serial passaging, and differentiation assays.
- Comparative transcriptomic analysis and drug testing with tyrosine kinase inhibitors (sorafenib, sunitinib).
Main Results:
- Successful generation of single-cell-derived tumorspheres with varying sphere formation rates for different cell lines.
- Expanded cells confirmed to possess stemness characteristics and distinct gene expression patterns compared to differentiated cells.
- Renal cancer stem cells demonstrated reduced sensitivity to sorafenib and sunitinib due to elevated tyrosine kinase expression.
Conclusions:
- Microfluidic single-cell culture arrays effectively expand renal cancer stem cells.
- This approach facilitates the drug testing of these rare cells.
- Findings highlight the potential for improved therapeutic strategies targeting renal cancer stem cells.
