Basic research for non-toxic cryopreservation of mammalian gamete without a use of membrane permeable cryoprotectants.

不使用膜渗透性冷冻保护剂的哺乳动物配子无毒冷冻保存的基础研究。

• 批准号: 15500302
• 负责人: KOSHIMOTO Chihiro
• 金额: $ 1.41万
• 依托单位: University of Miyazaki (2004)宮崎医科大学 (2003)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15500302/
• 关键词: cryopreservation; cryoprotectants; permeability; sperm; cooling rate; warming rate; strain differnces

Is addition of plasma membrane permeable cryoprotectants that believed to be required as a fundamental theory of cryobiology really essential for mammalian cell freezing? This is the very basic question to generate this research project. The objective of this project is to question the need for the addition of such kind of cell toxic chemicals for gamete cryopreservation.Our research plan was 1)Determination of physical parameters such as volume and membrane permeability, in mammalian sperm cells, which show complex in its morphology comparing to a common cell. 2)Identification of osmotic stress sensitivity of several rodent sperm to design new freezing extender without traditional cell-toxic cryoprotectants. 3)Establishing a protocol of sperm cryopreservation without traditional cryoprotectants for unique rodent except for mice and rats, according to the result of item 2). 4)Consideration of the effect of osmotic stress difference and cooling rate in mouse sperm freezability, in order … More to overcome strain differences of mouse sperm cryopreservation.The results are as follows ; 1)Possibility of sperm volume changes measurement in response to solution osmolality was suggested by use of flow cytometry, but uncompleted. 2)Other than mice and rats, osmotic stress sensitivities were also observed among 3 gerbils. 3)Cryopreserved Indian gerbil (Tatera indica) sperm without traditional cryoprotectants showed high survival after thawing, and in vitro fertilized Indian gerbil embryo developed to 2cell stage, (but only one example). 4)The strain difference of mouse sperm cryopreservation was suggested to be attributed to the difference of an optimum cooling rate (i.e.membrane permeability differences to water and chemicals) rather than the difference of cell sensitivity to osmotic stress.Our research has disclosed several problems that underlie the difficulties in freezability of rodent spermatozoa. Continuous work is required to lead a development of new protocol in gamete cryopreservation. Less

作为低温生物学的基本理论，加入质膜渗透性冷冻保护剂对哺乳动物细胞冷冻真的是必不可少的吗？这是产生这个研究项目的最基本的问题。本课题的研究目的是探讨在配子冷冻保存中是否需要添加这类细胞毒性物质。我们的研究计划是：1）测定哺乳动物精子细胞的体积和膜透性等物理参数。2)几种啮齿类动物精子渗透压敏感性的鉴定，以设计新的无细胞毒性冷冻保护剂的冷冻扩展剂。3)根据第2）项的结果，建立了除小鼠和大鼠外的独特啮齿类动物精子冷冻保存方案，不使用传统冷冻保护剂。4)考虑到渗透压差和降温速率对小鼠精子冷冻性的影响， ...更多信息 结果表明：1）用流式细胞仪测定精子体积随溶液渗透压的变化是可行的，但尚不完全。2)除小鼠和大鼠外，3只沙鼠也观察到渗透应激敏感性。3)印度沙鼠（Tatera indica）精子在不加传统冷冻保护剂的情况下冷冻保存，解冻后存活率高，体外受精胚胎发育至2-细胞期，但仅有一例。4)小鼠精子冷冻保存的品系差异是由于最佳冷却速率的差异（即细胞膜对水和化学物质的渗透性差异），而不是细胞对渗透压的敏感性差异。需要持续的工作来领导配子冷冻保存新方案的发展。少

项目成果

生命を凍らせて保存する-生物学的タイムマシーンにどこまで期待できるか-

通过冷冻来保存生命——我们对生物时间机器能有多少期望？

• 发表时间: 2003
• 期刊: Boundary 19
• 作者: Ichikawa;T. et al.;越本 知大
• 通讯作者: 越本 知大

Cryopreservation of Living Cell-What we can place our hope on the "biological time machine-.(in Japanese)

活细胞冷冻保存-我们可以对“生物时间机器”寄予什么希望-.（日语）

• 发表时间: 2003
• 期刊: Boundary 19
• 作者: Xiao N;Mamuti W;Yamasaki H;Sako Y;Nakao M;Nakaya K;Gottstein B;Schatz PM;Lihtowlers MW;Craig PS;Ito A.;Chihiro Koshimoto
• 通讯作者: Chihiro Koshimoto