Protocol for culture, drug treatment, and dissociation of patient-derived 3D ovarian cancer microspheroids for flow cytometry

Research Overview

Single-cell technologies reveal tumor heterogeneity and treatment response, but they need 3D cultures that can be taken apart gently enough to analyze. This protocol describes culturing patient-derived ovarian tumor cells as 3D microspheroids: preparing multi-well dishes, isolating tumor cells, generating and maintaining spheroids, then treating with drugs, recovering, and dissociating the spheroids for single-cell analysis by flow cytometry.

The workflow is scalable and cost-efficient and largely preserves the cellular composition of the original patient material — so drug-response readouts reflect the tumor as sampled rather than a selected subpopulation.