Protocol to optimize the biobanking of ovarian cancer organoids by accommodating patient-specific differences in stemness potential
Summary: 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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723004513 |
| _version_ | 1828718426489618432 |
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| author | Fabian Trillsch Bastian Czogalla Fabian Kraus Alexander Burges Sven Mahner Mirjana Kessler |
| author_facet | Fabian Trillsch Bastian Czogalla Fabian Kraus Alexander Burges Sven Mahner Mirjana Kessler |
| author_sort | Fabian Trillsch |
| collection | DOAJ |
| description | Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-12T14:43:33Z |
| format | Article |
| id | doaj.art-f9bb398ae3f54e9f8255e8cc3bb51e66 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-12T14:43:33Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-f9bb398ae3f54e9f8255e8cc3bb51e662023-08-16T04:27:19ZengElsevierSTAR Protocols2666-16672023-09-0143102484Protocol to optimize the biobanking of ovarian cancer organoids by accommodating patient-specific differences in stemness potentialFabian Trillsch0Bastian Czogalla1Fabian Kraus2Alexander Burges3Sven Mahner4Mirjana Kessler5Department of Obstetrics and Gynecology, University Hospital, LMU Munich, 81377 Munich, Germany; German Cancer Consortium (DKTK), Partner site Munich (LMU), 69120 Heidelberg, GermanyDepartment of Obstetrics and Gynecology, University Hospital, LMU Munich, 81377 Munich, GermanyDepartment of Obstetrics and Gynecology, University Hospital, LMU Munich, 81377 Munich, GermanyDepartment of Obstetrics and Gynecology, University Hospital, LMU Munich, 81377 Munich, GermanyDepartment of Obstetrics and Gynecology, University Hospital, LMU Munich, 81377 Munich, Germany; German Cancer Consortium (DKTK), Partner site Munich (LMU), 69120 Heidelberg, GermanyDepartment of Obstetrics and Gynecology, University Hospital, LMU Munich, 81377 Munich, Germany; German Cancer Consortium (DKTK), Partner site Munich (LMU), 69120 Heidelberg, Germany; Corresponding authorSummary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723004513Cell CultureCancerMicroscopyStem CellsOrganoids |
| spellingShingle | Fabian Trillsch Bastian Czogalla Fabian Kraus Alexander Burges Sven Mahner Mirjana Kessler Protocol to optimize the biobanking of ovarian cancer organoids by accommodating patient-specific differences in stemness potential STAR Protocols Cell Culture Cancer Microscopy Stem Cells Organoids |
| title | Protocol to optimize the biobanking of ovarian cancer organoids by accommodating patient-specific differences in stemness potential |
| title_full | Protocol to optimize the biobanking of ovarian cancer organoids by accommodating patient-specific differences in stemness potential |
| title_fullStr | Protocol to optimize the biobanking of ovarian cancer organoids by accommodating patient-specific differences in stemness potential |
| title_full_unstemmed | Protocol to optimize the biobanking of ovarian cancer organoids by accommodating patient-specific differences in stemness potential |
| title_short | Protocol to optimize the biobanking of ovarian cancer organoids by accommodating patient-specific differences in stemness potential |
| title_sort | protocol to optimize the biobanking of ovarian cancer organoids by accommodating patient specific differences in stemness potential |
| topic | Cell Culture Cancer Microscopy Stem Cells Organoids |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723004513 |
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