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
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.
