title: Biobanking of patient and patient-derived xenograft ovarian tumour tissue: efficient preservation with low and high fetal calf serum based methods.
abstract: Using patient-derived xenografts (PDXs) for preclinical cancer research demands proper storage of tumour material to facilitate logistics and to reduce the number of animals needed. We successfully established 45 subcutaneous ovarian cancer PDXs, reflecting all histological subtypes, with an overall take rate of 68%. Corresponding cells from mouse replaced human tumour stromal and endothelial cells in second generation PDXs as demonstrated with mouse-specific vimentin and CD31 immunohistochemical staining. For biobanking purposes two cryopreservation methods, a fetal calf serum (FCS)-based (95%v/v) "FCS/DMSO" protocol and a low serum-based (10%v/v) "vitrification" protocol were tested. After primary cryopreservation, tumour take rates were 38% and 67% using either the vitrification or FCS/DMSO-based cryopreservation protocol, respectively. Cryopreserved tumour tissue of established PDXs achieved take rates of 67% and 94%, respectively compared to 91% using fresh PDX tumour tissue. Genotyping analysis showed that no changes in copy number alterations were introduced by any of the biobanking methods. Our results indicate that both protocols can be used for biobanking of ovarian tumour and PDX tissues. However, FCS/DMSO-based cryopreservation is more successful. Moreover, primary engraftment of fresh patient-derived tumours in mice followed by freezing tissue of successfully established PDXs is the preferred way of efficient ovarian cancer PDX biobanking.
authors.affiliation: University of Groningen, University Medical Centre Groningen, Department of Gynaecologic Oncology, Groningen, The Netherlands.
authors.forename: Nicolette G
authors.initials: NG
authors.lastname: Alkema
authors.affiliation: University of Groningen, University Medical Centre Groningen, Department of Gynaecologic Oncology, Groningen, The Netherlands.
authors.forename: Tushar
authors.initials: T
authors.lastname: Tomar
authors.affiliation: University of Groningen, University Medical Centre Groningen, Department of Pathology and Medical Biology, Groningen, The Netherlands.
authors.forename: Evelien W
authors.initials: EW
authors.lastname: Duiker
authors.affiliation: University of Groningen, University Medical Centre Groningen, Department of Gynaecologic Oncology, Groningen, The Netherlands.
authors.forename: Gert
authors.initials: G
authors.lastname: Jan Meersma
authors.affiliation: University of Groningen, University Medical Centre Groningen, Department of Gynaecologic Oncology, Groningen, The Netherlands.
authors.forename: Harry
authors.initials: H
authors.lastname: Klip
authors.affiliation: University of Groningen, University Medical Centre Groningen, Department of Gynaecologic Oncology, Groningen, The Netherlands.
authors.forename: Ate G J
authors.initials: AG
authors.lastname: van der Zee
authors.affiliation: University of Groningen, University Medical Centre Groningen, Department of Gynaecologic Oncology, Groningen, The Netherlands.
authors.forename: G Bea A
authors.initials: GB
authors.lastname: Wisman
authors.affiliation: University of Groningen, University Medical Centre Groningen, Department of Medical Oncology, Cancer Research Centre Groningen, Groningen, The Netherlands.
authors.forename: Steven
authors.initials: S
authors.lastname: de Jong
mesh_terms: D000818:Animals; D018070:Biological Specimen Banks; D002417:Cattle; D018929:Cell Culture Techniques; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D005260:Female; D005312:Fetal Blood; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009368:Neoplasm Transplantation / Q000379:methods*; D010051:Ovarian Neoplasms / Q000473:pathology* / Q000601:surgery; D014022:Tissue Survival; D014183:Transplantation, Heterologous / Q000379:methods*; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents