Nutrition and Health Sciences, Department of


First Advisor

Janos Zempleni

Date of this Version

Summer 7-26-2021


Ogunnaike, M. (2021). Bovine Mammary Alveolar MAC-T Cells Afford a Tool for Studies of Bovine Milk Exosomes in Drug Delivery. Master’s thesis, University of Nebraska-Lincoln


A THESIS Presented to the Faculty of The Graduate College at the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Master of Science, Major: Nutrition, Under the Supervision of Professor Janos Zempleni. Lincoln, Nebraska: August, 2019

Copyright 2021 Ogunnaike Mojisola


Bovine milk exosomes (BMEs) have attracted attention as vehicles for delivering RNA therapeutics. BMEs originate in mammary alveolar cells. Here, we determined whether bovine mammary alveolar MAC-T cells are a promising tool to assess RNA delivery by BMEs. MAC-T cells exosomes (MAC-T BMEs) and BMEs were harvested by differential ultracentrifugation. Exosome size, morphology, microRNA content and marker proteins were assessed using nanoparticle tracking analysis, transmission electron microscopy, real-time PCR and immunoblot analysis, respectively. MAC-T cells were genetically engineered to secrete MAC-T BMEs endogenously labeled with a near-infrared fluorescent protein and tissue distribution was compared to fluorophore-labeled BMEs following intravenous injection in C57BL/6 mice. Morphology and size were similar in MAC-T BMEs and BMEs (94 ± 5.8 nm and 101 ± 4.2 nm, p > 0.05). Both preparations expressed miR-320a, miR-200c and let-7a-5p (positive controls) but not miR-1 (negative control). Exosome marker proteins, CD9, CD63, CD81 and Tsg101, were detected in both MAC-T BMEs and BMEs. The distribution in mouse tissues was similar for both preparations, with liver being the primary accumulation site. Collectively, MAC-T BMEs are a promising tool for BMEs-based RNA delivery studies.

Advisor: Janos Zempleni.