Novel cryoprotectant significantly improves the post-thaw recovery and quality of HSC from CB

Stylianou, J.; Vowels, M. R.; Hadfield, K.

doi:10.1080/14653240500501021

Cited by 36 publications

(34 citation statements)

References 9 publications

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“…There are two potential mechanisms for cell injury during freezing. 18,37 First, when water freezes in the cell suspending solution, solutes become more concentrated in the remaining liquid phase 21,37 that leads to cellular osmotic dehydration of the biological sample and injury/death. 38,39 Second, as cells freeze, intracellular ice crystals can form and directly puncture and damage the plasma membrane 18,19,24,39–41 , In order to prevent such freezing injury various cryoprotectants such as glycerol, ethanediol, propanediol and dimethyl sulfoxide (DMSO) are used for stable freezing and long-term preservation of established cell lines and tissues.…”

Section: Discussionmentioning

confidence: 99%

“…A method to facilitate the delayed use of previously generated early generation xenografts is cryopreservation. 18–21 This process entails freezing biological tissue for extended periods and is made possible with the use of cryoprotectant solutions that facilitate maintenance of cellular viability. Applying this method to PDX tumors allows for long-term storage of a living but frozen biobank of early generation PDX tumors with the ability to thaw and reimplant, or “reanimate”, the tumors at any future time point based on demand.…”

Section: Introductionmentioning

confidence: 99%

“…31 This product was designed to mitigate temperature induced cell stress responses by scavenging free radicals, pH buffering, provides oncotic/osmotic support, energy substrates, and ionic concentrations that balance the intracellular state at ultra-low temperature environments (−80°C to −196°C). 21,31–33 As this specialized media was specifically developed for the cryopreservation of extremely sensitive and vulnerable cell populations (human embryonic and pluripotent stem cells) we compared cryopreservation and subsequent reanimation engraftment outcomes between standard laboratory cryoprotectant media used in most PDX programs and this specialized media.…”

Section: Introductionmentioning

confidence: 99%

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

Ivanics

Bergquist

Liu

et al. 2018

Laboratory Investigation

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Introduction 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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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

Ivanics

Bergquist

Liu

et al. 2018

Laboratory Investigation

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“…The products are subjected to controlled rate freezing and then stored at very cold temperatures (below −150°C) until the units are required; at which time the products are thawed rapidly at 37°C, sometimes washed to remove the cryoprotectant DMSO, and administered to the patient [4–6, 11]. Unfortunately, these traditional cryopreservation media formulations and freezing methods introduce several distinct problems.…”

Section: Introductionmentioning

confidence: 99%

Improved post-thaw recovery of peripheral blood stem/progenitor cells using a novel intracellular-like cryopreservation solution

et al. 2009

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Background Peripheral blood stem cells (PBSC) have become the preferred stem cell source for autologous hematopoietic transplantation. A critical aspect of this treatment modality is cryopreservation of the stem cell products, which permits temporal separation of the PBSC mobilization/collection phase from the subsequent high-dose therapy. While controlled rate freezing and liquid nitrogen storage have become “routine” practice in many cell processing facilities, there is clearly room for improvement, as current cryopreservation media formulations still result in significant loss and damage to the stem/progenitor cell populations essential for engraftment, and can also expose the patients to relatively undefined serum components and larger volumes of DMSO that can contribute to the morbidity and mortality of the transplant therapy. Methods This study compared cryopreservation of PBSC in a novel intracellular-like, fully defined, serum- and protein-free preservation solution, CryoStor™ (BioLife Solutions, Inc.), with a standard formulation used by the Fred Hutchinson Cancer Research Center (FHCRC). Briefly, human PBSC apheresis specimens were collected and 5 × 107 cells/1 ml sample vial were prepared for cryopreservation in the following solutions: 1) FHCRC standard – Normosol-R, 5% HSA, 10% DMSO, and 2) CryoStor™ CS10 (final diluted conc. of 5% DMSO). A standard controlled-rate freezing program was employed, and frozen vials were stored in the vapor phase of a liquid nitrogen freezer for a minimum of one week. Vials were then thawed and evaluated for TNC, Viability, CD34, and granulocytes by flow cytometry, along with colony-forming activity in methylcellulose. Results The PBSC samples frozen in CryoStor™ CS10 yielded significantly improved post-thaw recoveries for total viable CD34+, CFU, and viable granulocytes. Specifically, relative to the FHCRC standard formulation, cryopreservation with CS10 resulted in an average 1.8 fold increased recovery of viable CD34+ cells (P = 0.005), a 1.5 fold increase in CFU-GM numbers (P = 0.030), and a 2.3 fold increase in granulocyte recovery (P = 0.045). Discussion This study indicates that use of CryoStor™ for cryopreservation can yield significantly improved recovery and in vitro functionality of the stem/progenitor cells in PBSC products. In addition, it is important to note that these improved recoveries were obtained while also not introducing any extra serum or serum-derived proteins, and reducing the final concentration/volume of DMSO by half. Further in vitro and in vivo studies are clearly necessary, however these findings imply use of CryoStor™ for cryopreservation might ultimately result in improved engraftment for those patients with lower content of CD34+ cells in their PBSC collections, along with reducing the requirement for additional apheresis collections, and decreasing the risk of adverse infusion reactions associated with higher exposure to DMSO.

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“…Complex cryopreservation media (CCM) including Viaspan, CryoStor, Unisol, Adesta, Celsior, and others, to name a few, when combined with CPAs for have been reported to improve cell survival to varying degrees. Improvements have been observed in systems including hepatocytes [ 21 , 22 ], cord blood stem cells [ 40 ], PBMC's [ 88 , 89 ], fi broblasts [ 20 ], keratinocytes [ 90 ], blood vessels [ 91 ] and engineered tissues [ 92 ]. In these studies, evaluation of the cryopreservation media was conducted and correlated with improvements in cell survival, function and growth.…”

Section: Carrier Mediamentioning

confidence: 99%

Biobanking: The Future of Cell Preservation Strategies

Baust

Corwin

VanBuskirk

2015

Advances in Experimental Medicine and Biology

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With established techniques cryopreservation is often viewed as an "old school" discipline yet modern cryopreservation is undergoing another scientific and technology development growth phase. In this regard, today's cryopreservation processes and cryopreserved products are found at the forefront of research in the areas of discovery science, stem cell research, diagnostic development and personalized medicine. As the utilization of cryopreserved cells continues to increase, the demands placed on the biobanking industry are increasing and evolving at an accelerated rate. No longer are samples providing for high immediate post-thaw viability adequate. Researchers are now requiring samples where not only is there high cell recovery but that the product recovered is physiologically and biochemically identical to its pre-freeze state at the genominic, proteomic, structural, functional and reproductive levels. Given this, biobanks are now facing the challenge of adapting strategies and protocols to address these needs moving forward. Recent studies have shown that the control and direction of the molecular response of cells to cryopreservation significantly impacts final outcome. This chapter provides an overview of the molecular stress responses of cells to cryopreservation, the impact of the apoptotic and necrotic cell death continuum and how studies focused on the targeted modulation of common and/or cell specific responses to freezing temperatures provide a path to improving sample quality and utility. This line of investigation has provided a new direction and molecular-based foundation guiding new research, technology development and procedures. As the use of and the knowledge base surrounding cryopreservation continues to expand, this path will continue to provide for improvements in overall efficacy and outcome.

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Novel cryoprotectant significantly improves the post-thaw recovery and quality of HSC from CB

Cited by 36 publications

References 9 publications

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

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

Improved post-thaw recovery of peripheral blood stem/progenitor cells using a novel intracellular-like cryopreservation solution

Biobanking: The Future of Cell Preservation Strategies

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