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确定人胎盘外植体培养和细胞外囊泡释放的最佳条件。

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

作者信息

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

机构信息

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.

出版信息

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

DOI:10.1016/j.isci.2023.108046
PMID:37829201
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10565782/
Abstract

Extracellular vesicles (EVs) can mediate intercellular communication, including signaling between the placenta and maternal tissues. Human placental explant culture is a versatile 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 modeling of short-term effects of environmental perturbations associated with pregnancy complications on placental function and EV release.

摘要

细胞外囊泡（EVs）可介导细胞间通讯，包括胎盘与母体组织之间的信号传递。人胎盘外植体培养是研究胎盘功能的一种通用模型系统。我们在不同的组织培养基类型和氧张力条件下进行了系统研究，以确定一种明确的无血清培养条件，该条件与含有可能被外源性EVs污染的培养基添加剂的传统未定义条件相比，能支持高滋养层细胞活力和代谢，以及释放相似群体的EVs。我们还确定了滋养层细胞活力和功能保持最佳的时间范围。使用经典的EV和胎盘特异性标志物的多重囊泡流式细胞术揭示了外植体来源的EVs的三个不同群体：小CD63 EVs；大PLAP+ EVs；以及CD63-/PLAP- EVs。这些培养和分析方法将有助于模拟与妊娠并发症相关的环境扰动对胎盘功能和EV释放的短期影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/cad7a199a793/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/85a56a775f3c/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/18e6f08226ef/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/617ef0ce52c8/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/1429959b5d93/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/82026e34dac5/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/621b7ccb7626/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/cad7a199a793/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/85a56a775f3c/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/18e6f08226ef/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/617ef0ce52c8/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/1429959b5d93/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/82026e34dac5/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/621b7ccb7626/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/10565782/cad7a199a793/gr6.jpg

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Generation and Application of a Reporter Cell Line for the Quantitative Screen of Extracellular Vesicle Release.用于细胞外囊泡释放定量筛选的报告细胞系的构建与应用
Front Pharmacol. 2021 Apr 16;12:668609. doi: 10.3389/fphar.2021.668609. eCollection 2021.
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Methamphetamine use alters human plasma extracellular vesicles and their microRNA cargo: An exploratory study.
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Placental small extracellular vesicles from normal pregnancy and gestational diabetes increase insulin gene transcription and content in β cells.正常妊娠和妊娠期糖尿病的胎盘小细胞外囊泡增加β 细胞中的胰岛素基因转录和含量。
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