Protocol for culture, drug treatment, and dissociation of patient-derived 3D ovarian cancer microspheroids for flow cytometry
Single-cell technologies provide insights into tumor heterogeneity and treatment response. Here, we present a protocol to culture patient-derived ovarian tumor cells as 3D microspheroids. We describe steps for preparing multi-well dishes and isolating ovarian tumor cells for spheroid generation and culture. We then detail procedures for drug treatment, recovery, and dissociation of spheroids followed by single-cell analysis via flow cytometry. This scalable and cost-efficient protocol largely preserves the cellular composition of the original material.
Authors
- Mariam Haffa
- Ulrika Joneborg (ORCID: https://orcid.org/0000-0002-1922-0738)
- Rita Ötvös
- Karolina Juhani (ORCID: https://orcid.org/0000-0002-9001-9447)
- Emelie Wallin (ORCID: https://orcid.org/0000-0002-0698-0930)
- Josefin Fernebro
- Olli-Pekka Kallioniemi
- Greta Gudoityte
- Brinton Seashore-Ludlow
Institutions
- Karolinska University Hospital (SE)
- Science for Life Laboratory (SE)
- Karolinska Institutet (SE)
- Institute for Molecular Medicine Finland (FI)
Publication Details
- Journal
- STAR Protocols
- Published
- 2026-03-31
- DOI
- https://doi.org/10.1016/j.xpro.2026.104480
- Primary Topic
- Single-cell and spatial transcriptomics
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Vetenskapsrådet
- Deutsche Krebshilfe
- Radiumhemmets Forskningsfonder
- Science for Life Laboratory