Flushed and Frozen: Developing Optimized Methods for Whole Body Perfusion of a Mouse Prior to Cryopreservation

School Name

Governor's School for Science & Mathematics

Grade Level

12th Grade

Presentation Topic

Physiology and Health

Presentation Type

Mentored

Mentor

Mentor: Demetri Spyropolous , Medical University of South Carolina

Written Paper Award

5th Place

Abstract

Every day, people are dying due to their inability to obtain organ transplants, as an organ will only remain viable for ten hours after it is harvested. A better method of organ preservation that does not compromise the tissue of the organ is required in order to extend the time from organ collection to transplant. The goal of this project is to develop a method for a whole body mouse perfusion that results in the most viable tissue after freezing and thawing back out. Tissue crystallization during in the freezing process varies in severity depending on how well the perfusate was distributed in the body, which results in nonviable tissue. We injected seven and a half blood volumes were injected into each mouse. Through four trials, a variety of perfusion methods were implemented, including use of different perfusion sites via the left and right ventricle, and different perfusates, such as PBS (phosphate buffered saline) and the cryoprotectant “Special K Special Sauce.” Blood samples were collected throughout the perfusion to measure the hematocrit levels as the perfusion progressed. The perfusion into the left ventricle using PBS was predicted to produce the most stable decline in hematocrit; however, the samples with “Special K Special Sauce" would be more viable after thawing, due to an antifreeze component that will prevent the tissue from crystalizing when cooled. The use of these hematocrit values as well as the future viability tests will provide insight into the best hope to develop an almost universal preservation method.

Location

Wall 318

Start Date

3-25-2017 9:00 AM

Presentation Format

Oral and Written

Group Project

No

COinS
 
Mar 25th, 9:00 AM

Flushed and Frozen: Developing Optimized Methods for Whole Body Perfusion of a Mouse Prior to Cryopreservation

Wall 318

Every day, people are dying due to their inability to obtain organ transplants, as an organ will only remain viable for ten hours after it is harvested. A better method of organ preservation that does not compromise the tissue of the organ is required in order to extend the time from organ collection to transplant. The goal of this project is to develop a method for a whole body mouse perfusion that results in the most viable tissue after freezing and thawing back out. Tissue crystallization during in the freezing process varies in severity depending on how well the perfusate was distributed in the body, which results in nonviable tissue. We injected seven and a half blood volumes were injected into each mouse. Through four trials, a variety of perfusion methods were implemented, including use of different perfusion sites via the left and right ventricle, and different perfusates, such as PBS (phosphate buffered saline) and the cryoprotectant “Special K Special Sauce.” Blood samples were collected throughout the perfusion to measure the hematocrit levels as the perfusion progressed. The perfusion into the left ventricle using PBS was predicted to produce the most stable decline in hematocrit; however, the samples with “Special K Special Sauce" would be more viable after thawing, due to an antifreeze component that will prevent the tissue from crystalizing when cooled. The use of these hematocrit values as well as the future viability tests will provide insight into the best hope to develop an almost universal preservation method.