Biobanking of patient and patient-derived xenograft ovarian tumour tissue: efficient preservation with low and high fetal calf serum based methods

被引:43
作者
Alkema, Nicolette G. [1 ]
Tomar, Tushar [1 ]
Duiker, Evelien W. [2 ]
Meersma, Gert Jan [1 ]
Klip, Harry [1 ]
van der Zee, Ate G. J. [1 ]
Wisman, G. Bea A. [1 ]
de Jong, Steven [3 ]
机构
[1] Univ Groningen, Univ Med Ctr Groningen, Dept Gynaecol Oncol, Groningen, Netherlands
[2] Univ Groningen, Univ Med Ctr Groningen, Dept Pathol & Med Biol, Groningen, Netherlands
[3] Univ Groningen, Univ Med Ctr Groningen, Canc Res Ctr Groningen, Dept Med Oncol, Groningen, Netherlands
关键词
CANCER XENOGRAFTS; IN-VITRO; GASTROINTESTINAL CANCER; CELL-LINE; MODELS; ESTABLISHMENT; VITRIFICATION; GROWTH; CARCINOMAS; MICE;
D O I
10.1038/srep14495
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];
摘要
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.
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页数:12
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