Protocol for generating and culturing high-grade serous ovarian carcinoma organoids from fresh or cryopreserved patient samples;
: Gall-Mas, Laura; Moyano-Galceran, Lidia; Valkonen, Elina; Hynninen, Johanna; Wennerberg, Krister; Senkowski, Wojciech
Publisher: Elsevier BV
: 2026
STAR protocols
: 104690
: 7
: 3
: 2666-1667
DOI: https://doi.org/10.1016/j.xpro.2026.104690
: https://doi.org/10.1016/j.xpro.2026.104690
: https://research.utu.fi/converis/portal/detail/Publication/526994911
Stable long-term high-grade serous ovarian carcinoma (HGSC) organoids can be difficult to develop due to limited primary sample growth in 3D conditions. Here, we present a detailed protocol for the development of stable HGSC organoids, including all steps from seeding primary cells through culture monitoring, passaging, and expansion to viable cryopreservation and resuscitation. We provide guidance on initial passaging, passaging ratios, and criteria for successful culture establishment. Moreover, we include a troubleshooting section tackling recurrent problems in HGSC organoid development.
cancer, cell culture, organoids
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This work was supported by the European Union’s Horizon 2020 research and innovation program under grant agreements no. 667403 for HERCULES (J. H. and K.W.); no. 965193 for DECIDER (J. H.); no. 845045 for RESIST3D (W.S.); and no. 101063359 for CROC (L.M.-G.); Danish Cancer Society, grant no. R204-A12322 (W.S.); Novo Nordisk Foundation: Interdisciplinary Synergy Programme 2021, grant no. NNF21OC0070381 (K.W.); Innovation Fund Denmark for ERA PerMed JTC2020 PARIS project, grant no. 0204-00005B (K.W.); and OvaCure Foundation (W.S., K.W., and J.H.). For all the EU-funded projects, views, and opinions expressed are those of the authors only and do not necessarily reflect those of the European Union or the European Research Executive Agency (REA). Neither the European Union nor the REA can be held responsible for them. The Graphical abstract/Figures were created using Biorender.com.
