【AI Explained】Establishment and differentiation of long-term trophoblast organoid cultures from the human placenta.

Literature Information

TL;DR

This study presents a novel three-dimensional organoid culture system for isolating and maintaining human trophoblast cells from first-trimester placentas, enabling long-term growth and differentiation into syncytiotrophoblast and extravillous trophoblast. This model closely mimics the architecture and function of the in vivo placenta, providing a valuable tool for investigating human placental development, function, and associated pregnancy disorders.

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Trophoblast · Organoid Culture · Human Placenta · Differentiation · Three-Dimensional Model

Abstract

The human placenta is essential for successful reproduction. There is great variation in the anatomy and development of the placenta in different species, meaning that animal models provide limited information about human placental development and function. Until recently, it has been impossible to isolate trophoblast cells from the human placenta that proliferate in vitro. This has limited our ability to understand pregnancy disorders. Generating an in vitro model that recapitulates the unique features of the human placenta has been challenging. The first in vitro model system of human trophoblast that could be cultured long term and differentiated to syncytiotrophoblast (SCT) and extravillous trophoblast (EVT) was a two-dimensional (2D) culture system of human trophoblast stem cells. Here, we describe a protocol to isolate trophoblast from first-trimester human placentas that can be grown long term in a three-dimensional (3D) organoid culture system. Trophoblast organoids can be established within 2-3 weeks, passaged every 7-10 d, and cultured for over a year. The structural organization of these human trophoblast organoids closely resembles the villous placenta with a layer of cytotrophoblast (VCT) that differentiates into superimposed SCT. Altering the composition of the medium leads to differentiation of the trophoblast organoids into HLA-G+ EVT cells which rapidly migrate and invade through the Matrigel droplet in which they are cultured. Our previous research confirmed that there is similarity between the trophoblast organoids and in vivo placentas in their transcriptomes and ability to produce placental hormones. This organoid culture system provides an experimental model to investigate human placental development and function as well as interactions of trophoblast cells with the local and systemic maternal environment.

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Primary Questions Addressed

1. What are the specific challenges in maintaining long-term trophoblast organoid cultures compared to traditional 2D cultures?
2. How does the differentiation of trophoblast organoids into SCT and EVT cells contribute to our understanding of placental function and pregnancy disorders?
3. In what ways can trophoblast organoid models be utilized to study maternal-fetal interactions and their implications for pregnancy health?
4. What potential applications do trophoblast organoids have in drug testing and development for pregnancy-related conditions?
5. How does the composition of the culture medium influence the functional characteristics of trophoblast organoids in vitro?

Key Findings

1. Research Background and Objective:

The human placenta plays a crucial role in successful reproduction, yet its complex anatomy and development vary significantly across species, rendering animal models inadequate for understanding human placental biology. Previous attempts to cultivate trophoblast cells—key components of the placenta—have faced significant challenges, particularly in producing viable long-term cultures that mimic the physiological properties of human placentas. This limitation has hindered research into pregnancy-related disorders. The objective of this study was to develop a long-term, three-dimensional (3D) organoid culture system to isolate and maintain trophoblast cells from human placentas, facilitating investigations into their development, differentiation, and interaction with maternal environments.

2. Main Methods and Findings:

The study established a protocol to isolate trophoblast cells from first-trimester human placentas, enabling their growth in a 3D organoid culture system. Within 2-3 weeks, trophoblast organoids could be established, passaged every 7-10 days, and cultured for over a year. The organoids exhibited structural organization that closely mimicked the in vivo villous placenta, featuring a layer of cytotrophoblast (VCT) that differentiated into syncytiotrophoblast (SCT). Notably, modification of the culture medium induced the differentiation of trophoblast organoids into HLA-G+ extravillous trophoblast (EVT) cells, which demonstrated rapid migration and invasion capabilities within the Matrigel environment. Transcriptomic analyses revealed that these organoids closely resembled in vivo placentas, particularly in their production of placental hormones.

3. Core Conclusion:

The successful establishment of long-term trophoblast organoid cultures represents a significant advancement in the field of reproductive biology. These organoids not only replicate the structural and functional characteristics of the human placenta but also provide a versatile platform for studying trophoblast biology, including cellular differentiation and their interactions with maternal tissues. This model opens new avenues for understanding placental development and the pathophysiology of pregnancy disorders.

4. Research Significance and Impact:

The development of a reliable 3D trophoblast organoid culture system has profound implications for both basic and clinical research. It offers a novel experimental model to investigate the complex mechanisms underlying human placentation and the role of trophoblasts in normal pregnancy and related disorders. This model can facilitate the identification of therapeutic targets for conditions such as preeclampsia, gestational diabetes, and miscarriage, ultimately contributing to improved maternal and fetal health outcomes. The insights gained from this organoid system could significantly enhance our understanding of human placental biology and its implications for reproductive health.

References

1. First-trimester determination of complications of late pregnancy. - Gordon C S Smith - JAMA (2010)
2. Development of the human placenta in the first three months of gestation. - W J HAMILTON;J D BOYD - Journal of anatomy (1960)
3. Development of the human placenta. - Margherita Y Turco;Ashley Moffett - Development (Cambridge, England) (2019)
4. Modeling human trophoblast, the placental epithelium at the maternal fetal interface. - Mariko Horii;Ojeni Touma;Tony Bui;Mana M Parast - Reproduction (Cambridge, England) (2020)
5. Derivation of Human Trophoblast Stem Cells. - Hiroaki Okae;Hidehiro Toh;Tetsuya Sato;Hitoshi Hiura;Sota Takahashi;Kenjiro Shirane;Yuka Kabayama;Mikita Suyama;Hiroyuki Sasaki;Takahiro Arima - Cell stem cell (2018)
6. Trophoblast organoids as a model for maternal-fetal interactions during human placentation. - Margherita Y Turco;Lucy Gardner;Richard G Kay;Russell S Hamilton;Malwina Prater;Michael S Hollinshead;Alasdair McWhinnie;Laura Esposito;Ridma Fernando;Helen Skelton;Frank Reimann;Fiona M Gribble;Andrew Sharkey;Steven G E Marsh;Stephen O’Rahilly;Myriam Hemberger;Graham J Burton;Ashley Moffett - Nature (2018)
7. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. - Toshiro Sato;Robert G Vries;Hugo J Snippert;Marc van de Wetering;Nick Barker;Daniel E Stange;Johan H van Es;Arie Abo;Pekka Kujala;Peter J Peters;Hans Clevers - Nature (2009)
8. Developmental differences in the expression of FGF receptors between human and mouse embryos. - T Kunath;Y Yamanaka;J Detmar;D MacPhee;I Caniggia;J Rossant;A Jurisicova - Placenta (2014)
9. Human chorionic gonadotrophin regulates FGF2 and other cytokines produced by human endometrial epithelial cells, providing a mechanism for enhancing endometrial receptivity. - P Paiva;N J Hannan;C Hincks;K L Meehan;E Pruysers;E Dimitriadis;L A Salamonsen - Human reproduction (Oxford, England) (2011)
10. Endometrial glands as a source of nutrients, growth factors and cytokines during the first trimester of human pregnancy: a morphological and immunohistochemical study. - Joanne Hempstock;Tereza Cindrova-Davies;Eric Jauniaux;Graham J Burton - Reproductive biology and endocrinology : RB&E (2004)

Literatures Citing This Work

1. Trophoblast organoids as a model for maternal-fetal interactions during human placentation. - Margherita Y Turco;Lucy Gardner;Richard G Kay;Russell S Hamilton;Malwina Prater;Michael S Hollinshead;Alasdair McWhinnie;Laura Esposito;Ridma Fernando;Helen Skelton;Frank Reimann;Fiona M Gribble;Andrew Sharkey;Steven G E Marsh;Stephen O’Rahilly;Myriam Hemberger;Graham J Burton;Ashley Moffett - Nature (2018)
2. Perinatal Derivatives: Where Do We Stand? A Roadmap of the Human Placenta and Consensus for Tissue and Cell Nomenclature. - Antonietta Rosa Silini;Roberta Di Pietro;Ingrid Lang-Olip;Francesco Alviano;Asmita Banerjee;Mariangela Basile;Veronika Borutinskaite;Günther Eissner;Alexandra Gellhaus;Bernd Giebel;Yong-Can Huang;Aleksandar Janev;Mateja Erdani Kreft;Nadja Kupper;Ana Clara Abadía-Molina;Enrique G Olivares;Assunta Pandolfi;Andrea Papait;Michela Pozzobon;Carmen Ruiz-Ruiz;Olga Soritau;Sergiu Susman;Dariusz Szukiewicz;Adelheid Weidinger;Susanne Wolbank;Berthold Huppertz;Ornella Parolini - Frontiers in bioengineering and biotechnology (2020)
3. Generation of Trophoblast-Like Cells From Hypomethylated Porcine Adult Dermal Fibroblasts. - Sharon Arcuri;Georgia Pennarossa;Fulvio Gandolfi;Tiziana A L Brevini - Frontiers in veterinary science (2021)
4. The Role of the 3Rs for Understanding and Modeling the Human Placenta. - Joana Costa;Ruth Mackay;Sophie-Christine de Aguiar Greca;Alessandro Corti;Elisabete Silva;Emmanouil Karteris;Arti Ahluwalia - Journal of clinical medicine (2021)
5. Mood Stabilizers in Psychiatric Disorders and Mechanisms Learnt from In Vitro Model Systems. - Ritu Nayak;Idan Rosh;Irina Kustanovich;Shani Stern - International journal of molecular sciences (2021)
6. Functional Analysis of p21Cip1/ and Its Family Members in Trophoblastic Cells of the Placenta and Its Roles in Preeclampsia. - Nina-Naomi Kreis;Alexandra Friemel;Lukas Jennewein;Samira Catharina Hoock;Anna Elisabeth Hentrich;Thorsten Nowak;Frank Louwen;Juping Yuan - Cells (2021)
7. Characterization of primary models of human trophoblast. - Megan A Sheridan;Xiaohui Zhao;Ridma C Fernando;Lucy Gardner;Vicente Perez-Garcia;Qian Li;Steven G E Marsh;Russell Hamilton;Ashley Moffett;Margherita Y Turco - Development (Cambridge, England) (2021)
8. Innate immune defenses at the maternal-fetal interface. - Eleanor C Semmes;Carolyn B Coyne - Current opinion in immunology (2022)
9. On Placental Toxicology Studies and Cerium Dioxide Nanoparticles. - Gaëlle Deval;Sonja Boland;Thierry Fournier;Ioana Ferecatu - International journal of molecular sciences (2021)
10. Modeling Innate Antiviral Immunity in Physiological Context. - Monty E Goldstein;Margaret A Scull - Journal of molecular biology (2022)

… (70 more literatures)

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