Technical Report

Patient-derived xenograft cryopreservation and reanimation outcomes are dependent on cryoprotectant type

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Patient-derived xenografts (PDX) are being increasingly utilized in preclinical oncologic research. Maintaining large colonies of early generation tumor-bearing mice is impractical and cost-prohibitive. Optimal methods for efficient long-term cryopreservation and subsequent reanimation of PDX tumors are critical to any viable PDX program. We sought to compare the performance of “Standard” and “Specialized” cryoprotectant media on various cryopreservation and reanimation outcomes in PDX tumors. Standard (10% DMSO media) and Specialized (Cryostor®) media were compared between overall and matched PDX tumors. Primary outcome was reanimation engraftment efficiency (REE). Secondary outcomes included time to tumor formation (TTF), time to harvest (TTH), and potential loss of unique PDX lines. Overall 57 unique PDX tumors underwent 484 reanimation engraftment attempts after previous cryopreservation. There were 10 unique PDX tumors cryopreserved with Standard (71 attempts), 40 with Specialized (272 attempts), and 7 with both (141 attempts). Median frozen time of reanimated tumors was 29 weeks (max. 177). Tumor pathology, original primary PDX growth rates, frozen storage times, and number of implantations per PDX model were similar between cryoprotectant groups. Specialized media resulted in superior REE (overall: 82 vs. 39%, p < 0.0001; matched: 97 vs. 36%, p < 0.0001; >52 weeks cryostorage: 59 vs. 9%, p < 0.0001), shorter TTF (overall 24 vs. 54 days, p = 0.0051; matched 18 vs. 53 days, p = 0.0013) and shorter TTH (overall: 64 vs. 89 days, p = 0.009; matched: 47 vs. 88 days, p = 0.0005) compared to Standard. Specialized media demonstrated improved REE with extended duration cryostorage (p = 0.048) compared to Standard. Potential loss of unique PDX lines was lower with Specialized media (9 vs. 35%, p = 0.017). In conclusion, cryopreservation with a specialized cryoprotectant appears superior to traditional laboratory-based media and can be performed with reliable reanimation even after extended cryostorage.

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The Mayo Clinic Robert D. and Patricia E. Kern Center for the Science of Health Care Delivery provides salary support for Dr. Ivanics and in kind material support for Dr. Bergquist. Dr. Bergquist receives salary support from the Mayo Clinic Clinician Investigator Training program. Dr. Fleming’s laboratory is funded by the Skip Viragh Foundation, the Various Donors in Pancreatic Cancer Research Fund, and the Research Animal Support Facility—Houston under NIH/NCI award (P30CA016672). Dr. Rios Perez was also supported in part by National Institutes of Health (NIH) grant T32CA009599. Dr. Truty is supported by Private Benefactor/Development,Departmental/Institutional Support, and Mayo Clinic/Karolinska Institute Grant.

Author information


  1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, Mayo Clinic Rochester, Rochester, MN, USA

    • Tommy Ivanics
    • , Gang Liu
    •  & Mark J Truty
  2. Robert D and Patricia E Kern Center for the Science of Healthcare Delivery, Mayo Clinic Rochester, Rochester, MN, USA

    • John R Bergquist
  3. Department of Surgical Oncology, University of Texas MD Anderson Cancer Center, Houston, TX, USA

    • Michael P. Kim
    • , Yaan Kang
    • , Matthew H. Katz
    • , Mayrim V. Rios Perez
    •  & Jason B Fleming
  4. Department of Surgery, University of Florida, Gainesville, FL, USA

    • Ryan M. Thomas
  5. Division of General Surgery, North Florida/South Georgia Veterans Health System, Gainesville, FL, USA

    • Ryan M. Thomas


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Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding author

Correspondence to Mark J Truty.