Identification of optimal conditions for human placental explant culture and extracellular vesicle release

Chandana Tekkatte; Scott A. Lindsay; Erika Duggan; Anelizze Castro-Martínez; Abbas Hakim; Isabella Saldana; Yan Zhang; Jun Zhou; Rachel Sebastian; Yukun Liu; Devin S. Pontigon; Morgan Meads; Tzu Ning Liu; Donald P. Pizzo; John Nolan; Mana M. Parast; Louise C. Laurent

iScience (Oct 2023)

Identification of optimal conditions for human placental explant culture and extracellular vesicle release

Chandana Tekkatte,
Scott A. Lindsay,
Erika Duggan,
Anelizze Castro-Martínez,
Abbas Hakim,
Isabella Saldana,
Yan Zhang,
Jun Zhou,
Rachel Sebastian,
Yukun Liu,
Devin S. Pontigon,
Morgan Meads,
Tzu Ning Liu,
Donald P. Pizzo,
John Nolan,
Mana M. Parast,
Louise C. Laurent

Affiliations

Chandana Tekkatte: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA; Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA
Scott A. Lindsay: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA; Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA
Erika Duggan: Scintillon Institute, San Diego, CA 92121, USA; Cellarcus Biosciences Inc, La Jolla, CA 92037, USA
Anelizze Castro-Martínez: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA; Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA
Abbas Hakim: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA; Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA
Isabella Saldana: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA; Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA
Yan Zhang: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA; Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA; Department of Obstetrics, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China
Jun Zhou: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA; Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA; Department of Obstetrics, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China
Rachel Sebastian: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA; Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA
Yukun Liu: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA; Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA; Department of Obstetrics and Gynecology, SUN Yat-sen Memorial Hospital, SUN Yat-sen University, Guangzhou, Guangdong, China
Devin S. Pontigon: Department of Pathology, University of California San Diego, La Jolla, CA 92037, USA
Morgan Meads: Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA; Department of Pathology, University of California San Diego, La Jolla, CA 92037, USA
Tzu Ning Liu: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA; Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA
Donald P. Pizzo: Department of Pathology, University of California San Diego, La Jolla, CA 92037, USA
John Nolan: Scintillon Institute, San Diego, CA 92121, USA; Cellarcus Biosciences Inc, La Jolla, CA 92037, USA
Mana M. Parast: Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA; Department of Pathology, University of California San Diego, La Jolla, CA 92037, USA
Louise C. Laurent: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA; Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA; Corresponding author

Journal volume & issue: Vol. 26, no. 10
p. 108046

Abstract

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Summary: Extracellular vesicles (EVs) can mediate intercellular communication, including signaling between the placenta and maternal tissues. Human placental explant culture is a versatile in vitro model system to investigate placental function. We performed systematic studies in different tissue culture media types and oxygen tensions to identify a defined serum-free culture condition that supports high trophoblast viability and metabolism, as well as the release of similar populations of EVs, compared to traditional undefined conditions that contain media additives potentially contaminated with exogenous EVs. We also determined the time frame in which trophoblast viability and functionality remain optimal. Multiplex vesicle flow cytometry with classical EV and placenta-specific markers revealed three separate populations of explant-derived EVs: small CD63+ EVs; large PLAP+ EVs; and CD63-/PLAP- EVs. These culture and analytical approaches will enable in vitro modeling of short-term effects of environmental perturbations associated with pregnancy complications on placental function and EV release.

Published in iScience

ISSN: 2589-0042 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science
Website: http://www.cell.com/iscience/home

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