Maintaining viability and characteristics of cholangiocarcinoma tissue by vitrification-based cryopreservation

Zeng, Min; Yang, Qiu-Rui; Fu, Gongbo; Zhang, Yuan; Zhou, Xu; Huang, Weijian; Zhang, Hong-Dan; Li, Weijian; Wang, Zhenyu; Yan, He‐Xin; Zhai, Bo

doi:10.1016/j.cryobiol.2017.07.004

Cited by 5 publications

(4 citation statements)

References 27 publications

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“…After a period of cryopreservation, cholangiocarcinoma demonstrated the highest rate (70%) of successful secondary PDX generation. This result is similar to that reported by Zeng et al wherein cryopreservation techniques for cholangiocarcinoma were analyzed [25] . We also demonstrated similar secondary engraftment salvage (60%) for pancreatic cancers, another malignant tumor type of which we have our largest proportion in our PDX catalog.…”

Section: Discussionsupporting

confidence: 91%

Successful Secondary Engraftment of Pancreatic Ductal Adenocarcinoma and Cholangiocarcinoma Patient-Derived Xenografts After Previous Failed Primary Engraftment

Hernandez

Yang

Leiting

et al. 2019

Translational Oncology

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BACKGROUND: Patient-derived xenografts (PDX) provide histologically accurate cancer models that recapitulate patient malignant phenotype and allow for highly correlative oncologic in-vivo downstream translational studies. Primary PDX engraftment failure has significant negative consequences on programmatic efficiency and resource utilization and is due to either no tumor growth or development of lymphoproliferative tumors. We aimed to determine if secondary engraftment of previously cryopreserved patient tumor tissues would allow salvage of PDX models that failed previous primary engraftment and increase overall engraftment efficiency. METHODS: Patient hepatobiliary and pancreatic cancers that failed primary engraftment were identified. Previously cryopreserved primary patient cancerous tissues were implanted into immunodeficient mice (NOD/SCID). Mice were monitored, growth metrics calculated, and secondary engraftment outcomes were recorded. Established PDX were verified and compared to original patient tissue through multiple generations by a GI pathologist. RESULTS: We identified 55 patient tumors that previously failed primary engraftment: no tumor growth (n = 46, 84%) or lymphoproliferative tumor (LT) (n = 9, 16%). After secondary implantation using cryopreserved patient tissues, 29 new histologically validated PDX models were generated with an overall secondary engraftment rate of 53% for all tumor types with greatest yield in pancreatic and biliary tract cancers. Of the secondary engraftment failures (n = 26), 21 (38%) were due to no growth and 5 (9%) developed LT. CONCLUSION: Secondary PDX engraftment using cryopreserved primary cancerous is feasible after previous failed engraftment attempts and can result in a 50% increase in overall engraftment efficiency with decreases in LT formation. This technique allows for salvage of critical patient PDX models that would otherwise not exist. SYNOPSIS: Patient-derived xenografts have many important translational applications however can be limited by engraftment failure. We demonstrate optimized methodology utilizing cryopreservation of primary tumor tissue that allows for subsequent successful secondary engraftment and creation of PDX models that failed previous primary engraftment and allowed salvage of patient PDX models that would otherwise not exist.

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Section: Discussionsupporting

confidence: 91%

Successful Secondary Engraftment of Pancreatic Ductal Adenocarcinoma and Cholangiocarcinoma Patient-Derived Xenografts After Previous Failed Primary Engraftment

Hernandez

Yang

Leiting

et al. 2019

Translational Oncology

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“…Notably, this method avoided ice crystallization, by which the cell viability may be reduced (21). Furthermore, the required time and basic cost of this cryopreservation method were lower than those of conventional cryopreservation methods (22).…”

Section: Discussionmentioning

confidence: 98%

Cryopreserved biopsy tissues of rectal cancer liver metastasis for assessment of anticancer drug response in�vitro and in�vivo

et al. 2019

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Living tumors are of great scientific value for clinical medicine and basic research, especially for drug testing. An increasing number of drug tests fail due to the use of imperfect models. The aim of the present study was to develop a novel method combining vitrification-based cryopreservation of tumor biopsies and precision-cut slice cultivation for the assessment of anticancer drug responses. Biological characteristics of rectal cancer liver metastasis biopsies could be retained by vitrification-based cryopreservation. The patient-derived xenograft models were successfully established using both fresh and warmed biopsy tissues. Precision-cut slicing provided a similar three-dimensional architecture and heterogeneity to the original tumor. The positive drug responses in the xenograft model were consistent with those in precision-cut slice cultures in vitro. The present study demonstrated that live tumor biopsies could be preserved using vitrification-based cryopreservation. The warmed tissues developed xenograft tumors, which were also useful for either in vivo or in vitro anticancer drug testing. Precision-cut slices derived from the warmed tissues provided an efficient tool to assess anticancer drug response in vitro.

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“…Therefore, a reliable and efficient cryopreservation method for living tissue is indispensable. Vitrification-based cryopreservation method can be developed to preserve fresh tissue, by which the biological characteristics of the original tumor can be retained and the utilization of specimens may be markedly improved ( 11 ).…”

Section: Introductionmentioning

confidence: 99%

A pre‑clinical model combining cryopreservation technique with precision‑cut slice culture method to assess the in vitro drug response of hepatocellular carcinoma

et al. 2022

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Models considering hepatocellular carcinoma (Hcc) complexity cannot be accurately replicated in routine cell lines or animal models. We aimed to evaluate the practicality of tissue slice culture by combining it with a cryopreservation technique. We prepared 0.3-mm-thick tissue slices by a microtome and maintained their cell viability using a cryopreservation technique. Slices were cultured individually in the presence or absence of regorafenib (REG) for 72 h. Alterations in morphology and gene expression were assessed by histological and genetic analysis. Overall viability was also analyzed in tissue slices by CCK-8 quantification assay and fluorescent staining. Tissue morphology and cell viability were evaluated to quantify drug effects. Histological and genetic analyses showed that no significant alterations in morphology and gene expression were induced by the vitrification-based cryopreservation method. The viability of warmed Hcc tissues was up to 90% of the fresh tissues. The viability and proliferation could be retained for at least four days in the filter culture system. The positive drug responses in precision-cut slice culture in vitro were evaluated by tissue morphology and cell viability. In summary, the successful application of precision-cut Hcc slice culture combined with a cryopreservation technique in a systematic drug screening demonstrates the feasibility and utility of slice culture method for assessing drug response.

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Maintaining viability and characteristics of cholangiocarcinoma tissue by vitrification-based cryopreservation

Cited by 5 publications

References 27 publications

Successful Secondary Engraftment of Pancreatic Ductal Adenocarcinoma and Cholangiocarcinoma Patient-Derived Xenografts After Previous Failed Primary Engraftment

Successful Secondary Engraftment of Pancreatic Ductal Adenocarcinoma and Cholangiocarcinoma Patient-Derived Xenografts After Previous Failed Primary Engraftment

Cryopreserved biopsy tissues of rectal cancer liver metastasis for assessment of anticancer drug response in�vitro and in�vivo

A pre‑clinical model combining cryopreservation technique with precision‑cut slice culture method to assess the in vitro drug response of hepatocellular carcinoma

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