TY - JOUR
T1 - Bovine mammary alveolar MAC-T cells afford a tool for studies of bovine milk exosomes in drug delivery
AU - Ogunnaike, Mojisola
AU - Wang, Haichuan
AU - Zempleni, Janos
N1 - Funding Information:
The authors acknowledge the use of the Biomedical and Obesity Research Core (BORC) in the Nebraska Center for the Prevention of Obesity Disease through Dietary Molecules at the University of Nebraska-Lincoln. The BORC receives support from NIH (NIGMS) IDeA award 2P20GM104320. This work was supported by the National Institutes of Health [grant numbers 1P20GM104320]; the National Institute of Food and Agriculture [grant numbers 2016-67001-25301, 2020-67017-30834, 2021-99999-99999]; and the United States Department of Agriculture [grant numbers Hatch-1011996, W4002] (to J.Z).
Funding Information:
The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Janos Zempleni reports financial support was provided by National Institutes of Health. Janos Zempleni reports financial support was provided by National Institute of Food and Agriculture. Janos Zempleni reports was provided by United States Department of Agriculture. Janos Zempleni reports a relationship with PureTech Health, Inc. that includes: board membership. Janos Zempleni reports a relationship with University of Nebraska-Lincoln that includes: non-financial support. Janos Zempleni reports a relationship with The Japanese Society for Food Factors that includes: speaking and lecture fees and travel reimbursement. Janos Zempleni has patent pending to PureTech Health, Inc.].
Funding Information:
This work was supported by the National Institutes of Health [grant numbers 1P20GM104320]; the National Institute of Food and Agriculture [grant numbers 2016-67001-25301, 2020-67017-30834, 2021-99999-99999]; and the United States Department of Agriculture [grant numbers Hatch-1011996, W4002] (to J.Z).
Funding Information:
The authors acknowledge the use of the Biomedical and Obesity Research Core (BORC) in the Nebraska Center for the Prevention of Obesity Disease through Dietary Molecules at the University of Nebraska-Lincoln. The BORC receives support from NIH (NIGMS) IDeA award 2P20GM104320.
Publisher Copyright:
© 2021 Elsevier B.V.
PY - 2021/12/15
Y1 - 2021/12/15
N2 - 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 afford a 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. Distribution in mouse tissues was similar for both preparations, with liver being the primary accumulation site. Collectively, MAC-T BMEs afford a tool for BMEs-based RNA delivery studies.
AB - 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 afford a 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. Distribution in mouse tissues was similar for both preparations, with liver being the primary accumulation site. Collectively, MAC-T BMEs afford a tool for BMEs-based RNA delivery studies.
KW - Bovine mammary alveolar MAC-T cells
KW - Bovine milk exosomes
KW - CD81 fusion protein
KW - Drug delivery
KW - RNA
KW - Transgenic
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UR - http://www.scopus.com/inward/citedby.url?scp=85118721085&partnerID=8YFLogxK
U2 - 10.1016/j.ijpharm.2021.121263
DO - 10.1016/j.ijpharm.2021.121263
M3 - Article
C2 - 34742829
AN - SCOPUS:85118721085
SN - 0378-5173
VL - 610
JO - International journal of pharmaceutics
JF - International journal of pharmaceutics
M1 - 121263
ER -