Minimizing the role of cryoprotectant toxicity for cryopreservation

最大限度地减少冷冻保护剂毒性对冷冻保存的作用

• 批准号: 8925076
• 负责人: Utkan Demirci
• 金额: $ 38.94万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8925076
• 关键词: Animals; Area; Biodiversity; Biological Assay; Biological Preservation; Blood; Blood Banks; Caring; Cell Volumes; Cells; Clinic; Clinical; Couples; Cryopreservation; Devices; Diffusion; Excision; Fertility; Freezing; Germ Cells; Goals; Hepatocyte; Human; Ice; In Vitro; Infertility; Knowledge; Lead; Life; Manuals; Mechanics; Membrane; Methods; Microfluidics; Oocytes; Osmotic Shocks; Outcome; Outcome Study; Process; Publishing; Regenerative Medicine; Reproductive Medicine; Role; Sampling; Stem cells; System; Technology; Tissue Engineering; Tissues; Toxic effect; Training; Variant; Work; base; cell injury; cell type; clinically significant; cytotoxic; drug testing; improved; islet; nanolitre; new technology; novel; novel strategies; sperm cell; transplantation medicine

DESCRIPTION (provided by applicant): Long-term biopreservation of cells and tissues has a broad impact in multiple fields including tissue engineering, regenerative medicine, stem cells, blood banking, animal strain preservation (biodiversity protection), clinical sample storage, transplantation medicine and in vitro drug testing. Vitrification (ice/crystal-free cryopreservatio) has emerged as a novel approach over traditional slow freezing methods. Although vitrification minimizes mechanical damage due to ice crystal nucleation, it suffers from toxicity due to high concentrations of cryoprotectant agents (CPAs). The current vitrification methods require extremely high levels of CPAs of up to 8.2 M that are cytotoxic and cause osmotic shock. Also, the lengthy manual processing steps of current vitrification methods add to the technical complexity, require highly trained technicians, and result in variations between users. For instance, low CPA-level vitrification has immense potential for the stem cells compared to other methods in preserving their functionality. Recently, we demonstrated that we can achieve vitrification at ultra-rapid freezing and thawing rates with as low as 1.5M CPA concentration. We are adapting this new knowledge to the vital needs of cell cryopreservation at the clinic including discarded anonymous human oocytes. This proposal investigates a new experimental strategy to minimize the CPA concentrations and improve clinical outcomes using novel technologies (i.e., nanoliter droplet vitrification). These steps are facilitated by theoretical understanding o the underlying mechanisms governing vitrification. The expected outcome of this study is a closed-system platform technology with broad applications to human cell (e.g., hepatocytes, oocytes, sperm, stem cells), tissues (e.g., blood), micro-tissues (e.g., embryoid bodies, islets) covering areas of reproductive medicine, tissue engineering and regenerative medicine as well as to wild life preservation. These studies can also significantly impact the care of infertile couples and facilitate fertility preservation.

描述（由申请人提供）：细胞和组织的长期生物保存在多个领域具有广泛的影响，包括组织工程、再生医学、干细胞、血库、动物品系保存（生物多样性保护）、临床样本储存、移植医学和体外药物测试。玻璃化（无冰/晶体冷冻保存）已成为一种优于传统缓慢冷冻方法的新颖方法。尽管玻璃化可以最大程度地减少冰晶成核造成的机械损伤，但它会因高浓度的冷冻保护剂 (CPA) 而产生毒性。目前的玻璃化方法需要高达 8.2 M 的极高水平的 CPA，这些 CPA 具有细胞毒性并导致渗透压休克。此外，当前玻璃化方法冗长的手动处理步骤增加了技术复杂性，需要训练有素的技术人员，并导致用户之间存在差异。例如，与其他方法相比，低 CPA 水平的玻璃化冷冻对于干细胞在保留其功能方面具有巨大的潜力。最近，我们证明我们可以在低至 1.5M CPA 浓度的情况下以超快速冷冻和解冻速率实现玻璃化。我们正在将这些新知识应用于临床细胞冷冻保存的重要需求，包括 丢弃的匿名人类卵母细胞。该提案研究了一种新的实验策略，以最大限度地减少 CPA 浓度并使用新技术（即纳升液滴玻璃化）改善临床结果。对玻璃化的基本机制的理论理解促进了这些步骤。这项研究的预期成果是一种封闭系统平台技术，可广泛应用于人体细胞（如肝细胞、卵母细胞、精子、干细胞）、组织（如血液）、微组织（如胚状体、胰岛）涵盖生殖医学、组织工程和再生医学以及野生动物保护领域。这些研究还可以显着影响不孕夫妇的护理并促进生育力的保存。

项目成果

Bio-inspired solute enables preservation of human oocytes using minimum volume vitrification.

• DOI: 10.1002/term.2439
• 发表时间: 2018-01
• 期刊: Journal of tissue engineering and regenerative medicine
• 影响因子: 3.3
• 作者: Choi JK;El Assal R;Ng N;Ginsburg E;Maas RL;Anchan RM;Demirci U
• 通讯作者: Demirci U

Multiscale assembly for tissue engineering and regenerative medicine.

• DOI: 10.1016/j.tibtech.2015.02.003
• 发表时间: 2015-05
• 期刊: TRENDS IN BIOTECHNOLOGY
• 影响因子: 17.3
• 作者: Guven, Sinan;Chen, Pu;Inci, Fatih;Tasoglu, Saves;Erkmen, Burcu;Demirci, Utkan
• 通讯作者: Demirci, Utkan

Preserving human cells for regenerative, reproductive, and transfusion medicine.

• DOI: 10.1002/biot.201300074
• 发表时间: 2014-07
• 期刊: BIOTECHNOLOGY JOURNAL
• 影响因子: 4.7
• 作者: Asghar, Waseem;El Assal, Rami;Shafiee, Hadi;Anchan, Raymond M.;Demirci, Utkan
• 通讯作者: Demirci, Utkan

Engineering cancer microenvironments for in vitro 3-D tumor models.

• DOI: 10.1016/j.mattod.2015.05.002
• 发表时间: 2015-12
• 期刊: Materials today (Kidlington, England)
• 作者: Asghar W;El Assal R;Shafiee H;Pitteri S;Paulmurugan R;Demirci U
• 通讯作者: Demirci U

Levitational Image Cytometry with Temporal Resolution.

• DOI: 10.1002/adma.201405660
• 发表时间: 2015-07-08
• 期刊: Advanced materials (Deerfield Beach, Fla.)
• 作者: Tasoglu S;Khoory JA;Tekin HC;Thomas C;Karnoub AE;Ghiran IC;Demirci U
• 通讯作者: Demirci U