Basic research for non-toxic cryopreservation of mammalian gamete without a use of membrane permeable cryoprotectants.
| Project/Area Number |
15500302
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Laboratory animal science
|
|---|
| Research Institution | University of Miyazaki (2004)
宮崎医科大学 (2003) |
|---|
Principal Investigator |
KOSHIMOTO Chihiro University of Miyazaki, Research Center for Frontier Biosciences, Associate Professor, フロンティア科学実験総合センター, 助教授 (70295210)
|
|---|
| Project Period (FY) |
2003 – 2004
|
| Project Status |
Completed (Fiscal Year 2004)
|
| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2004: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2003: ¥1,700,000 (Direct Cost: ¥1,700,000)
|
|---|
| Keywords | cryopreservation / cryoprotectants / permeability / sperm / cooling rate / warming rate / strain differnces |
|---|
| Research Abstract |
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
|
Report
(3 results)
Research Products
(2 results)
