Date on Senior Honors Thesis

5-2021

Document Type

Senior Honors Thesis

Degree Name

B.A.

Department

Biology

Degree Program

College of Arts and Sciences

Author's Keywords

red blood cells; cryopreservation; trehalose; hemolysis; biomimetics; cell preservation

Abstract

One of the most common medical procedures performed in US hospitals is blood transfusions. Unfortunately, the red blood cells (RBCs) for transfusion have a limited shelf life after donation due to detrimental storage effects on morphological and biochemical properties. Inspired by nature, I am developing a biomimetics approach to preserve RBCs for long-term storage using compounds that occur in animals that have developed a natural propensity to survive in a frozen or desiccated state for decades. Trehalose was employed as a cryoprotective agent when added to the extracellular freezing solution. The highest percent of RBCs with intact membranes after freezing and thawing was obtained using a cryopreservation solution comprised of 250 million RBCs/mL, 300 mM trehalose, 100 mM NaCl, in 20 mM HEPES buffer, pH 7.1. Under these conditions RBCs with intact membranes were recovered at 86 ± 12%. I have demonstrated the effectiveness and feasibility of using trehalose as a cryoprotective agent, and morphological intact RBCs were recovered after freezing and thawing with low cellular loss.

Lay Summary

One of the most common medical procedures performed in US hospitals is blood transfusions. Unfortunately, the red blood cells (RBCs) for transfusion have a limited shelf life after donation due to damage to the cells that occurs because of storage. Inspired by nature, we are developing an approach to preserve RBCs for long-term storage using compounds that occur in animals that have developed a strategy to survive in a frozen or desiccated state for decades. The sugar, trehalose was employed as a compound to protect against freezing induced damage (cryoprotective agent) when added to a solution containing RBCs. The percent of RBCs with intact membranes after freezing and thawing was close to 100% when trehalose was present, and no cells were recovered in the absence of trehalose. We have demonstrated the effectiveness and feasibility of using trehalose as a cryoprotective agent and RBCs were recovered after freezing and thawing with low cellular loss.

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