Date of Graduation

12-2020

Document Type

Thesis

Degree Name

Bachelor of Science in Agricultural, Food and Life Sciences

Department

Animal Science

Advisor/Mentor

Huang, Yan

Committee Member/Reader

Rosenkrans, Charles

Committee Member/Second Reader

Kegley, Beth

Abstract

Stem cell research opens a wide range of possibilities from lab-grown tissues for medical purposes to cloned animals for livestock production. The issue with one of the most popular forms of cell recombination into a stem cell, known as somatic cell nuclear transfer (SCNT), is that it requires an oocyte. The requirement of an oocyte is often the host of public scrutiny for its economic and ethical issues. Platelets may offer a less expensive and more accessible alternative to oocytes as sources of naturally enucleated cells. Thus, the objective of this explorative research project is to isolate and fuse platelets to form a large body of cytoplasm, which could be used to receive a donated somatic nucleus for recombination. Blood samples were collected first from a horse and then from cattle. The whole blood was centrifuged to create a density gradient, and then the platelets were isolated using a commercially available protocol (Abcam, Cambridge, MA) for the isolation of human platelets. Once isolated, the platelets were diluted using an RPMI solution to four dilutions using the volumetric ratio of ml of platelet-rich plasma (PRP) to ml of RPMI solution. The dilutions were 1:10, 1:100, 1:1000, and 1:10,000. Two samples of each dilution were then treated with either a 25%, 50%, or 100% concentration of polyethylene glycol (PEG). Fusion was found in the 1:100 dilution sample treated with the 100% PEG concentration. The rest of the samples appeared to have been destroyed. Two further replications were inconclusive as the platelets did not survive the isolation process. Any future attempts were canceled due to the shutdowns related to the COVID-19 pandemic. Despite inconclusive results, the fusion found in the first trial is encouraging that future research might find a method to consistently obtain platelet fusion.

Keywords

Platelets, isolation, fusion, recombination

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