Microencapsulation of oocytes for low-CPA (cryoprotectant) vitrification

用于低 CPA（冷冻保护剂）玻璃化冷冻的卵母细胞微囊化

基本信息

批准号：
8600270
负责人：
Xiaoming He
金额：
$ 33万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-03-01 至 2016-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8600270
关键词：
Affect Alginates Benchmarking Biological Models Biological Preservation Breeding Caliber Cell Survival Cells Cryopreservation Dehydration Development Devices Effectiveness Electrostatics Embryo Encapsulated Equilibrium Exposure to Fertility Fertilization in Vitro Freezing Future Goals Human Ice In Vitro Injury Kinetics Life Liquid substance Livestock Maintenance Mammalian Cell Mammals Mechanics Medical Metabolic Methods Microcapsules drug delivery system Microencapsulations Modeling Oocytes Outcome Performance Peromyscus Phase Transition Procedures Protocols documentation Public Health Quartz Radiosurgery Reproductive Medicine Research Solutions Stress Techniques Testing Time Water Woman Work animal resource cell injury chemotherapy gonad function improved in vivo nanolitre new technology novel novel strategies occupational hazard public health relevance

项目摘要

DESCRIPTION (provided by applicant): Oocyte cryopreservation is of great importance to the advancement of assisted reproductive medicine, maintenance of animal resources, and livestock management. However, the commonly used methods today for oocyte cryopreservation by either slow-freezing or conventional vitrification have inherent drawbacks. For example, the slow-freezing (freezing: the transition of liquid water into ice crystal) approach is associated with inevitable cell injury due to ice formation and slow- freezing induced cell dehydration. The unusually high CPA (cryoprotectant) concentration (4 - 7 M) required by the conventional vitrification (vitrification: the transition of liquid water into an amorphous, glassy state rather than ice crystal) method can result in significant metabolic and osmotic injury in living cells even in a short exposure time of only a few minutes. Presumably, it is these inherent drawbacks that are responsible for the dismal outcome of oocyte cryopreservation to date. The goal of the proposed research outlined in this R01 proposal is to develop new strategies for cell cryopreservation by microencapsulating the cells in alginate microcapsule to vitrify at a low-CPA (low and non- toxic amount of cryoprotectants, = 1.5 M) concentration. The proposed low-CPA vitrification approach combines all the advantages of the commonly used slow-freezing and conventional vitrification techniques today while avoiding all their shortcomings. Oocytes of the naturally bred (outbred) Peromyscus will be used as the biological model in this project so that the results obtained from the proposed studies can be more transferable to achieve low-CPA vitrification of oocytes of other naturally bred mammals including humans. In addition, Peromyscus embryos will be used as the benchmark biological model in this project to test the new approach in view of the fact that embryo cryopreservation has been successful in general. It is believed that the proposed research and the novel low-CPA vitrification approach will have a significant impact on the field of oocyte cryopreservation for assisted reproductive medicine, maintenance of animal resources, and livestock management. PUBLIC HEALTH RELEVANCE: Oocyte cryopreservation is of great importance to the advancement of assisted reproductive medicine. We propose to develop a novel technology to achieve much improved performance for oocyte cryopreservation. This research will have a significant impact on the preservation of future fertility of women who may lose gonadal function because of exposure to environmental/occupational hazards or aggressive medical treatments such as extirpative surgery, radiation, and chemotherapy.

描述（申请人提供）：卵母细胞冷冻保存对于辅助生殖医学的进步、动物资源的维护和牲畜管理具有重要意义。然而，当今常用的卵母细胞冷冻保存方法（缓慢冷冻或传统玻璃化冷冻）具有固有的缺点。例如，缓慢冷冻（冷冻：液态水转变为冰晶）方法与由于冰形成和缓慢冷冻引起的细胞脱水而导致的不可避免的细胞损伤有关。传统玻璃化（玻璃化：液态水转变为无定形、玻璃态而不是冰晶）方法所需的异常高的 CPA（冷冻保护剂）浓度（4 - 7 M）可能会导致活细胞显着的代谢和渗透损伤即使只有几分钟的短暂曝光时间。据推测，正是这些固有的缺点导致了迄今为止卵母细胞冷冻保存的惨淡结果。 R01 提案中概述的研究目标是通过将细胞封装在藻酸盐微胶囊中，以低 CPA（低且无毒量的冷冻保护剂 = 1.5 M）浓度进行玻璃化，从而开发细胞冷冻保存的新策略。所提出的低 CPA 玻璃化方法结合了当今常用的慢速冷冻和传统玻璃化技术的所有优点，同时避免了它们的所有缺点。本项目将使用自然繁殖（远交）Peromyscus 的卵母细胞作为生物学模型，以便从拟议的研究中获得的结果可以更好地转移，以实现包括人类在内的其他自然繁殖哺乳动物的卵母细胞的低 CPA 玻璃化冷冻。此外，鉴于胚胎冷冻保存总体上已取得成功，本项目将使用Peromyscus胚胎作为基准生物模型来测试新方法。据信，所提出的研究和新型低CPA玻璃化冷冻方法将对辅助生殖医学的卵母细胞冷冻保存、动物资源维护和牲畜管理领域产生重大影响。公共健康相关性：卵母细胞冷冻保存对于辅助生殖医学的进步非常重要。我们建议开发一种新技术来显着提高卵母细胞冷冻保存的性能。这项研究将对因暴露于环境/职业危害或积极的医疗治疗（如摘除手术、放射和化疗）而可能丧失性腺功能的女性未来生育能力的保护产生重大影响。

项目成果

期刊论文数量（35）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

In vitro culture of early secondary preantral follicles in hanging drop of ovarian cell-conditioned medium to obtain MII oocytes from outbred deer mice.

在卵巢细胞条件培养基悬滴中体外培养早期次级腔前卵泡，以获得远交鹿小鼠的 MII 卵母细胞。

DOI：
发表时间：
2013-12
期刊：
影响因子：
0
作者：
Choi, Jung Kyu;Agarwal, Pranay;He, Xiaoming
通讯作者：
He, Xiaoming

Alginate Hydrogel Microencapsulation Inhibits Devitrification and Enables Large-Volume Low-CPA Cell Vitrification.

海藻酸盐水凝胶微胶囊可抑制失透并实现大容量低 CPA 细胞玻璃化。

DOI：
发表时间：
2015-11-25
期刊：
影响因子：
19
作者：
Huang, Haishui;Choi, Jung Kyu;Rao, Wei;Zhao, Shuting;Agarwal, Pranay;Zhao, Gang;He, Xiaoming
通讯作者：
He, Xiaoming

A Biomimetic Core-Shell Platform for Miniaturized 3D Cell and Tissue Engineering.

用于微型 3D 细胞和组织工程的仿生核壳平台。

DOI：
发表时间：
2015-08
期刊：
影响因子：
0
作者：
Agarwal, Pranay;Choi, Jung Kyu;Huang, Haishui;Zhao, Shuting;Dumbleton, Jenna;Li, Jianrong;He, Xiaoming
通讯作者：
He, Xiaoming

Bioengineering of injectable encapsulated aggregates of pluripotent stem cells for therapy of myocardial infarction.

用于治疗心肌梗塞的可注射封装多能干细胞聚集体的生物工程。

DOI：
发表时间：
2016-10-27
期刊：
影响因子：
16.6
作者：
Zhao, Shuting;Xu, Zhaobin;Wang, Hai;Reese, Benjamin E;Gushchina, Liubov V;Jiang, Meng;Agarwal, Pranay;Xu, Jiangsheng;Zhang, Mingjun;Shen, Rulong;Liu, Zhenguo;Weisleder, Noah;He, Xiaoming
通讯作者：
He, Xiaoming

The Unusual Properties of Polytetrafluoroethylene Enable Massive-Volume Vitrification of Stem Cells with Low-Concentration Cryoprotectants.

聚四氟乙烯的独特特性使得能够使用低浓度冷冻保护剂对干细胞进行大体积玻璃化冷冻。

DOI：
发表时间：
2019-01
期刊：
影响因子：
6.8
作者：
Cao, Yuan;Zhao, Gang;Panhwar, Fazil;Zhang, Xiaozhang;Chen, Zhongrong;Cheng, Lin;Zang, Chuanbao;Liu, Feng;Zhao, Yuanjin;He, Xiaoming
通讯作者：
He, Xiaoming