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. 2023 Sep 15;4(3):102484.
doi: 10.1016/j.xpro.2023.102484. Epub 2023 Aug 15.

Protocol to optimize the biobanking of ovarian cancer organoids by accommodating patient-specific differences in stemness potential

Fabian Trillsch  1 , Bastian Czogalla  2 , Fabian Kraus  2 , Alexander Burges  2 , Sven Mahner  1 , Mirjana Kessler  3
Affiliations

Protocol to optimize the biobanking of ovarian cancer organoids by accommodating patient-specific differences in stemness potential

Fabian Trillsch et al. STAR Protoc. .

Abstract

We present a protocol for effective biobanking of epithelial ovarian cancer organoids, considering the heterogeneous clinical presentation and high recurrence rates. Our protocol involves parallel testing of three media to identify patient-specific optimal conditions. We describe steps for tissue dissociation, differential seeding, organoid cultivation, and biobanking. We outline procedures for fixation, embedding, and staining for confocal imaging. Furthermore, we demonstrate that brief cultivation of isolates in 2D on plastic enhances organoid-forming potential in selected lines, expanding their application scope. For complete details on the use and execution of this protocol, please refer to Hoffmann et al.1.

Keywords: Cancer; Cell Culture; Microscopy; Organoids; Stem Cells.

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Conflict of interest statement

Declaration of interests M.K. is listed as an inventor on a patent related to a medium for ovarian cancer organoids. F.T. received research funding, honoraria, and travel expenses from and is a member of the advisory board of AstraZeneca, Clovis, Eisai, ImmunoGen, Medac, MSD, PharmaMar, Roche, and Tesaro/GSK. S.M. received research funding, honorary, or travel expenses from and is a member of the advisory board of AbbVie, AstraZeneca, Clovis, Eisai, GlaxoSmithKline, Hubro, Medac, MSD, Novartis, Nykode, Olympus, PharmaMar, Pfizer, Roche, Sensor Kinesis, Teva, and Tesaro.

Figures

None
Graphical abstract
Figure 1
Figure 1
Preparation of the fixed organoids for the paraffin embedding
Figure 2
Figure 2
Example of High-grade serous (HGS) ovarian cancer line, the most frequent histological EOC type, where the brief expansion of epithelial progenitors in 2D positively influenced organoid growth in subsequent 3D culture. Scale bar=500 µm.
Figure 3
Figure 3
Examples of 3 donor tissues with diverging outcomes in different media, with the best result observed in OCM1, OCM1+HRG-B1 and OCM2 from left to right respectively. Scale bar= 500µm.
Figure 4
Figure 4
Illustration of the sustained long-term growth potential of the EOC organoid lines. Scale bar=200µm.
Figure 5
Figure 5
Confocal images of parental tumor tissue and organoid line, showing ubiquitous expression of SOX17 in tumor compartment and intact epithelial architecture (CDH1). Scale bar=20 µm
Figure 6
Figure 6
Example of slow-growing organoid line, showing vital growth pattern for 1 month after seeding. Scale bar= 500µm
See this image and copyright information in PMC

References

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