1.Phenotypic plasticity and secretory heterogeneity in subpopulations derived from single cancer cell.
Zhun LIN ; Siping LIANG ; Zhe PU ; Zhengyu ZOU ; Luxuan HE ; Christopher J LYON ; Yuanqing ZHANG ; Tony Y HU ; Minhao WU
Acta Pharmaceutica Sinica B 2025;15(5):2723-2735
Single-cell analysis of phenotypic plasticity could improve the development of more effective therapeutics. Still, the development of tools to measure single-cell heterogeneity has lagged due to difficulties in manipulating and culturing single cells. Here, we describe a single-cell culture and phenotyping platform that employs a starburst microfluidic network and automatic liquid handling system to capture single cells for long-term culture and multi-dimensional analysis and quantify their clonal properties via their surface biomarker and secreted cytokine/growth factor profiles. Studies performed on this platform found that cells derived from single-cell cultures maintained phenotypic equilibria similar to their parental populations. Single-cell cultures exposed to chemotherapeutic drugs stochastically disrupted this balance to favor stem-like cells. They had enhanced expression of mRNAs and secreted factors associated with cell signaling, survival, and differentiation. This single-cell analysis approach can be extended to analyze more complex phenotypes and screen responses to therapeutic targets.
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