Feasibility of large-scale oocyte cryobanking of C57BL/6N mice and the influence of donor microbiome on recovery and genomic stability

C57BL/6N 小鼠大规模卵母细胞冷冻银行的可行性以及供体微生物组对恢复和基因组稳定性的影响

• 批准号: 10610678
• 负责人: CRAIG L FRANKLIN
• 金额: $ 45.33万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10610678
• 关键词: Anaphase; Aneuploidy; Animals; Archives; Breeding; C57BL/6 Mouse; C57BL/6N Mouse; Charge; Chromosome abnormality; Chromosomes; Communities; Competence; Cryopreservation; Data; Derivation procedure; Development; Disease model; Embryo; Embryo Transfer; Embryonic Development; Equus caballus; Estrus; Evaluation; Event; Female; Fertilization in Vitro; Follicle Stimulating Hormone; Generations; Genetic; Genetic Drift; Genetically Engineered Mouse; Genome; Genome Stability; Genomics; Germ Cells; Goals; Gonadotropins; Haploidy; In Vitro; Live Birth; Logistics; Metabolic; Metagenomics; Metaphase; Methods; Microtubules; Missouri; Mitochondrial DNA; Modeling; Mouse Strains; Mus; Mutant Strains Mice; Oocytes; Outcome; Phenotype; Physiological; Play; Population; Pregnancy loss; Procedures; Production; Publications; Recovery; Research; Research Personnel; Resources; Resuscitation; Rivers; Role; Savings; Serum; Shipping; Ships; Sperm Banks; Superovulation; Surrogate Mothers; Testing; Time; Transgenic Mice; Universities; Variant; Vendor; Weather; blastocyst; cost; cryobiology; embryo cryopreservation; embryo quality; experience; experimental study; genetic strain; genome sequencing; gut microbiota; human error; improved; in vivo; inhibin; meetings; microbial; microbiome; mouse model; oocyte cryopreservation; oocyte quality; operation; preservation; pup; response; segregation; sperm cell; sperm cryopreservation; success; tool; transcriptomics; whole genome

Project Summary The goal of this project is to establish large-scale oocyte cryopreserve and assess the viability, cost and animal savings, as well as determine what factors may influence the recovery of viable rederived mice. Our experience in cryopreservation has shown varying quality of recovered oocytes among different vendors, even within a specific mouse strain. We will cryopreserve oocytes from C57BL/6N mice from the four most commonly used vendors (Jackson, Charles River, Taconic and Envigo). We compare the quality of oocytes through IVF to the blastocyst stage as well as to live birth. We will also examine possible corollaries to embryo quality including genome aneuploidy and the microbiome of the donor dams. This data will inform our implementation of large-scale preservation and rederivation and identify factors that influence successful outcomes. This proposal will lead to efficiencies in cryopreservation of haploid gametes (oocytes) and success would; i) reduce costs related animal shipments required on demand of a project ii) would eliminate the requirement to synchronize oocyte production and ET recipient mice’s estrus cycle, iii) enable testing of each batch of oocytes for developmental competency, iv) improve successful live pup production rates and reduce the time between when a researcher orders an IVF recovery and we can provide animals, v) and collectively, the use of cryopreserved oocytes will reduce the operating expenses of MMRRC. This information will be shared with the other three MMRRCs either during our monthly consortium meetings or during our annual consortium meeting. Additionally, this information will be shared widely through publications and presentations.

项目摘要 本课题的目的是建立大规模的卵母细胞冻存库， 成本和动物储蓄，以及确定哪些因素可能影响恢复可行的 再衍生小鼠我们在冷冻保存方面的经验表明， 卵母细胞之间的不同供应商，甚至在一个特定的小鼠品系。我们会冷冻保存 来自四个最常用的供应商（杰克逊，Charles River、Taconic和Envigo）。我们比较了体外受精卵母细胞和囊胚的质量， 阶段以及活产。我们还将研究胚胎质量的可能推论，包括 供体母鼠的基因组非整倍性和微生物组。这些数据将告知我们的 实施大规模保护和再衍生，并确定影响因素 成功的结果。这一建议将导致单倍体的超低温保存的效率 配子（卵母细胞）和成功将; i）减少所需的动物运输成本， 项目的需求ii）将消除同步卵母细胞生产和ET的要求 受体小鼠的发情周期，iii）能够测试每批卵母细胞的发育 能力，iv）提高成功的活仔生产率，并减少 一个研究人员订购试管婴儿恢复，我们可以提供动物，v）和集体，使用 冷冻保存的卵母细胞将减少MMRRC的运营费用。此信息将 与其他三个MMRR在我们的月度联盟会议或 我们的年度联盟会议此外，这些信息将通过以下方式广泛共享： 出版物和介绍。