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Storage of Extracellular Vesicles in Human Milk, and MicroRNA Profiles in Human Milk Exosomes and Infant Formulas

Leiferman, Amy*; Shu, Jiang; Upadhyaya, Bijaya*; Cui, Juan; Zempleni, Janos*

Journal of Pediatric Gastroenterology and Nutrition: May 6, 2019 - Volume Publish Ahead of Print - Issue - p
doi: 10.1097/MPG.0000000000002363
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ABSTRACT The objectives of this study were to lay the methodological groundwork for field studies of microRNA analysis in exosomes from small sample volumes of human milk, and assess exosome and microRNA content in infant formulas. When human milk was stored at 4°C for four weeks, the count of exosome-sized vesicles decreased progressively to 49% ± 13% of that in fresh milk. Exosomes were purified from 1 mL of fresh human milk and their microRNA content was assessed by microRNA-sequencing analysis and compared to that in infant formulas. We identified 221 microRNAs in exosomes from three samples of fresh human milk; 84 microRNAs were present in all three samples. MicroRNAs were not detectable in infant formulas and their exosome-sized vesicles, which appeared to be casein micelles. We conclude that large scale studies of microRNAs in human milk exosomes are feasible, and exosomes and microRNAs are not detectable in formulas.

*Department of Nutrition & Health Sciences, University of Nebraska-Lincoln

Department of Computer Science & Engineering, University of Nebraska-Lincoln.

Address correspondence and reprint requests to Janos Zempleni, 316C Leverton Hall, Lincoln, NE 68583-0806, USA (e-mail: jzempleni2@unl.edu).

Received 4 January, 2019

Accepted 23 March, 2019

Source of Funding: JZ receives research support from the National Institute of Food and Agriculture (NIFA), U.S. Department of Agriculture, under award numbers 2015-67017-23181 and NIFA2016-67001-25301, National Institutes of Health (NIH) grant 1P20GM104320, the University of Nebraska Agricultural Research Division (Hatch Act), U.S. Department of Agriculture multistate group W3002, and the University of Nebraska President's Office. JC receives research support from the University of Nebraska President's Office. JZ serves as a consultant for PureTech Health, Inc.

Conflicts of Interest: None declared.

Authors Roles: Amy Leiferman acquired data, made substantial contributions to the interpretation of data, drafted and revised this work, gave final approval of the work to be published and agreed to be accountable for all aspects of the work. Jiang Shu acquired data, made substantial contributions to the interpretation of data, drafted and revised this work, gave final approval of the work to be published and agreed to be accountable for all aspects of the work. Bijaya Upadhyaya acquired data, made substantial contributions to the interpretation of data, drafted and revised this work, gave final approval of the work to be published and agreed to be accountable for all aspects of the work. Juan Cui acquired data, made substantial contributions to the interpretation of data, drafted and revised this work, gave final approval of the work to be published and agreed to be accountable for all aspects of the work. Janos Zempleni conceived the idea for this work, made substantial contributions to the interpretation of data, drafted and revised this work, gave final approval of the work to be published and agreed to be accountable for all aspects of the work.

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© 2019 by European Society for Pediatric Gastroenterology, Hepatology, and Nutrition and North American Society for Pediatric Gastroenterology,