Selected Grantee Publications
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- 11 results found
- Preservation
Establishing the Hybrid Rat Diversity Program: A Resource for Dissecting Complex Traits
Dwinell et al., Mammalian Genome. 2025.
https://pubmed.ncbi.nlm.nih.gov/39907792
Rat models have been extensively used for studying human complex disease mechanisms, behavioral phenotypes, and environmental factors and for discovering and developing drugs. Systems genetics approaches have been used to study the effects of both genetic variation and environmental factors. This approach recognizes the complexity of common disorders and uses intermediate phenotypes to find relationships between genetic variation and clinical traits. This article describes the Hybrid Rat Diversity Program (HDRP) at the Medical College of Wisconsin, which involves 96 inbred rat strains and aims to provide a renewable and reusable resource in terms of the HRDP panel of inbred rat strains, the genomic data derived from the HRDP strains, and banked resources available for additional studies. Supported by ORIP (R24OD024617) and NHLBI.
Establishment of a Practical Sperm Cryopreservation Pathway for the Axolotl (Ambystoma mexicanum): A Community-Level Approach to Germplasm Repository Development
Coxe et al., Animals (Basel). 2024.
https://pubmed.ncbi.nlm.nih.gov/38254376/
The axolotl (Ambystoma mexicanum) is an important biomedical research model for organ regeneration, but housing and maintaining live animals is expensive and risky as new transgenic lines are developed. The authors report an initial practical pathway for sperm cryopreservation to support germplasm repository development. They assembled a pathway through the investigation of axolotl sperm collection by stripping, refrigerated storage in various osmotic pressures, cryopreservation in various cryoprotectants, and in vitro fertilization using thawed sperm. This work is the first report of successful production of axolotl offspring with cryopreserved sperm and provides a general framework for pathway development to establish Ambystoma germplasm repositories for future research and applications. Supported by ORIP (R24OD010441, R24OD028443, P40OD019794).
Conduction-Dominated Cryomesh for Organism Vitrification
Guo et al., Advanced Science. 2024.
https://pubmed.ncbi.nlm.nih.gov/38018294/
Vitrification-based cryopreservation via cryomesh is a promising approach for maintaining biodiversity, health care, and sustainable food production via long-term preservation of biological systems. Here, researchers conducted a series of experiments aimed at optimizing the cooling and rewarming rates of cryomesh to increase the viability of various cryopreserved biosystems. They found that vitrification was significantly improved by increasing thermal conductivity, reducing mesh wire diameter and pore size, and minimizing the nitrogen vapor barrier of the conduction-dominated cryomesh. Cooling rates increased twofold to tenfold in a variety of biosystems. The conduction-dominated cryomesh improved the cryopreservation outcomes of coral larvae, Drosophila embryos, and zebrafish embryos by vitrification. These findings suggest that the conduction-dominated cryomesh can improve vitrification in such biosystems for biorepositories, agriculture and aquaculture, and research. Supported by ORIP (R24OD028444, R21OD028758, R24OD034063, R21OD028214), NIDDK, and NIGMS.
Simian Immunodeficiency Virus and Storage Buffer: Field-Friendly Preservation Methods for RNA Viral Detection in Primate Feces
Wilde et al., mSphere. 2023.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10732032/
Simian immunodeficiency virus (SIV) infects more than 40 nonhuman primate (NHP) species in sub-Saharan Africa, but testing in wild NHP populations can be challenging. Researchers compared methods for SIV RNA preservation and recovery from NHP fecal samples stored in four different buffers. The goal of this work was to identify a robust “field-friendly” method (i.e., without freezing or refrigeration) for this effort, and the samples were collected from a mantled guereza colobus housed at the Columbus Zoo and Aquarium. The authors reported that the DNA/RNA shield is an optimal buffer for preserving SIV RNA in fecal samples in the field. Their findings will inform future fieldwork and facilitate improved approaches for studies of SIV and other RNA viruses. Supported by ORIP (P51OD011132) and NIAID.
Zebrafish as a High Throughput Model for Organ Preservation and Transplantation Research
Da Silveira Cavalcante et al., The FASEB Journal. 2023.
https://faseb.onlinelibrary.wiley.com/doi/10.1096/fj.202300076R
Organ transplantation increases the quality of life and life expectancy of patients with chronic end-stage diseases, but the preservation of organs for transplantation remains a significant barrier. In the current study, researchers demonstrate the value of zebrafish as a high-throughput model organism in the fields of solid-organ preservation and transplantation, with a focus on heart preservation via partial freezing. Their techniques have the potential to advance research in the fields of cryobiology and solid-organ transplantation. Supported by ORIP (R24OD031955) and NHLBI.
Orthotopic Transplantation of the Full-Length Porcine Intestine After Normothermic Machine Perfusion
Abraham et al., Transplantation Direct. 2022.
https://www.doi.org/10.1097/TXD.0000000000001390
Successful intestinal transplantation currently is hindered by graft injury that occurs during procurement and storage, which contributes to postoperative sepsis and allograft rejection. Improved graft preservation could expand transplantable graft numbers and enhance post-transplant outcomes. Superior transplant outcomes recently have been demonstrated in clinical trials using machine perfusion to preserve the liver. The investigators report the development and optimization of machine perfusion preservation of small intestine and successful transplantation of intestinal allografts in a porcine model. Supported by ORIP (K01OD019911), NIAID, and NIDDK.
Rapid Joule Heating Improves Vitrification Based Cryopreservation
Zhan et al., Nature Communications. 2022.
https://www.doi.org/10.1038/s41467-022-33546-9
Cryopreservation by vitrification is an effective approach for long-term preservation of biosystems, but effective vitrification often requires high concentrations of cryoprotective agent (CPA), which can be toxic. The investigators described a joule heating–based platform technology for rapid rewarming of biosystems, which allows the use of low concentrations of CPA. They demonstrated the success of this platform in cryopreservation of three model systems: adherent cells, Drosophila melanogaster embryos, and rat kidney slices with low CPA concentrations. This work provides a general solution to cryopreserve a broad spectrum of cells, tissues, organs, and organisms. Supported by ORIP (R21OD028758), NIDDK, NHLBI, and NIGMS.
Nonhuman Primate Models for SARS-CoV-2 Research: Cryopreservation as a Means to Maintain Critical Models and Enhance the Genetic Diversity of Colonies
Arnegard and Hild et al., Lab Animal. 2021.
https://doi.org/10.1038/s41684-021-00792-1
This commentary, written by ORIP staff, addresses the need for improved cryopreservation methods and resources for nonhuman primate (NHP) gametes and embryos to safeguard newly developed NHP models and enhance the genetic diversity of NHP colonies without reliance on animal importations. Cryopreservation also plays critical roles in medical approaches to preserve the fertility of patients who must undergo potentially gonadotoxic treatments, as well as nascent genome editing efforts to develop new NHP models for human diseases. Given these diverse benefits to research progress, ORIP continues to fund the development of cryopreservation tools and approaches for NHPs and other animal models.
Cryopreservation and Preparation of Thawed Spermatozoa from Rhesus Macaques (Macaca mulatta) for In Vitro Fertilization
De Carvalho et al., Journal of the American Association for Laboratory Animal Science. 2021.
https://www.ingentaconnect.com/content/aalas/jaalas/pre-prints/content-jaalas-20-000028
Optimizing procedures for cryopreservation and subsequent thawing for rhesus macaques is required to prevent cryodamage that negatively impacts artificial insemination and in vitro fertilization rates. Investigators systematically assessed two cryopreservation methods and four recovery methods in three interdependent experiments. Results demonstrated that slow-freezing, coupled with density gradient centrifugation provided the highest efficacy in functional sperm for in vitro use. Additional studies are required to further optimize sperm cryopreservation in rhesus macaques. Supported by ORIP (P51OD011092).
Cryopreservation Method for Drosophila melanogaster Embryos
Zhan et al., Nature Communications. 2021.
https://www.nature.com/articles/s41467-021-22694-z
Drosophila melanogaster is a premier model for biomedical research. However, preservation of Drosophila stocks is labor intensive and costly. Researchers at University of Minnesota reported an efficient method for cryopreservation by optimizing key steps including embryo permeabilization and cryoprotectant agent loading. This method resulted in more than 10% of embryos developing into fertile adults after cryopreservation for 25 distinct strains from different sources. The further optimization and wide adoption of this protocol will solve the long-standing issue in reliably preserving Drosophila stocks and will significantly impact Drosophila as a model organism for biomedical research. Supported by ORIP (R21OD028758) and NIGMS.