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

doi:10.1016/j.isci.2023.108046

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

iScience. 2023 Sep 26;26(10):108046. doi: 10.1016/j.isci.2023.108046. eCollection 2023 Oct 20.

Authors

Chandana Tekkatte^{1

2}, Scott A Lindsay^{1

2}, Erika Duggan^{3

4}, Anelizze Castro-Martínez^{1

2}, Abbas Hakim^{1

2}, Isabella Saldana^{1

2}, Yan Zhang^{1

2

5}, Jun Zhou^{1

2

5}, Rachel Sebastian^{1

2}, Yukun Liu^{1

2

6}, Devin S Pontigon⁷, Morgan Meads^{2

7}, Tzu Ning Liu^{1

2}, Donald P Pizzo⁷, John Nolan^{3

4}, Mana M Parast^{2

7}, Louise C Laurent^{1

2}

Affiliations

¹ Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Diego, La Jolla, CA 92037, USA.
² Sanford Consortium for Regenerative Medicine, La Jolla, CA 92037, USA.
³ Scintillon Institute, San Diego, CA 92121, USA.
⁴ Cellarcus Biosciences Inc, La Jolla, CA 92037, USA.
⁵ Department of Obstetrics, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.
⁶ Department of Obstetrics and Gynecology, SUN Yat-sen Memorial Hospital, SUN Yat-sen University, Guangzhou, Guangdong, China.
⁷ Department of Pathology, University of California San Diego, La Jolla, CA 92037, USA.

Abstract

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.

Keywords: Biological sciences; Cell biology; Stem cell research.

Abstract

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