Trophoblasts, 1st ed. 2024 Methods and Protocols Methods in Molecular Biology Series, Vol. 2728

This volume explores the latest approaches used to assess trophoblast angiogenesis, transport function, cellular respirations, migration, and invasion. The chapters in this book cover topics such as various methods to study and manipulate primary trophoblast cells; strategies using liposomes to deliver biomolecules to trophoblasts; in vitro models of the placenta that emulate the cellular interactions in the 3D uterine environment; and models exploring heterogenous cell types and 3D-organoid structures that represent cell-to-cell interactions in vivo. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and thorough, Trophoblasts: Methods and Protocols is a valuable resource that will help researchers better understand the function of the placenta and its complex contribution to maternal health and fetal development, and also provide greater insights to the area of study known as the developmental origins of health and diseases (DoHaD). 

Preface…

Table of Contents…

Contributing Authors…

Part I Isolation and Culture Methods

1. Isolation and Maintenance in Culture of Primary Human Trophoblast from term Placentae

Yohanes N. S. Nursalim, Katie M. Groom, Cherie Blenkiron, and Lawrence W. Chamley

2. Isolation of Human Villous Cytotrophoblastic Cells from Term Placenta for Transfection

Caroline Toudic and Benoit Barbeau

3. Enzymatic Digestion and Single Cell Isolation of Peri-Implantation Stage Human Trophoblast Cells

Deirdre M. Logsdon, Toshihko Ezashi, and Ye Yuan

4. Precision-Cut Slice Culture Method for Rat Placenta

Fusun Gundogan, Jeffrey Gilligan, and Suzanne de la Monte

Part II Analysis of Fusion

5. Evaluation of Molecular Interactions and Cellular Dynamics at the Maternal-Fetal Interface during Placental Morphogenesis

Madhurima Paul and Rupasri Ain

6. In Vivo Quantitative Assessment of Gestational Choriocarcinoma Development and Progression using Luminescent Trophoblast Cells

Wael Traboulsi, Deborah Reynaud, Roland Abi Nahed, Frédéric Sergent, Nadia Alfaidy, and Mohamed Benharouga

7. Analyzing Trophoblast Fusion using Immunofluorescence and Split Protein Complementation Assays

Adam Jaremek and Stephen J. Renaud

8. Quantification of Trophoblast Syncytialization by Fluorescent membrane Labeling

Yang Zhang and Huanghe Yang

9. Assessment of Placental Sodium Independent Leucine Uptake and Transfer in Trophoblast Cells

Jonas Zaugg and Christiane Albrecht

10. Assessing Cholesterol Efflux on Primary Human Trophoblast Cells

Barbara Fuenzalida and Christiane Albrecht

11. RGD-Based Fluorescence to Assess Placental Angiogenesis

Veronique Josserand, Jonathan Lavaud, Michelle Keramidas, Constance Collet, Wael Traboulsi, Pascale Hoffmann, Jean-Jacques Feige, Mohamed Benharouga, Jean-Luc Coll, and Nadia Alfaidy

12. Real-Time Assessment of Mitochondrial Function in Cytotrophoblast and Syncytialized Trophoblast Cells using the Seahorse XFe24 Extracellular Flux Analyzer

O’Llenecia S. Walker, Linda L. May, and Sandeep Raha

13. Quantifying Trophoblast Function using the xCELLigence System

Rosemary J. Keogh and Padma Murthi

Part III Targeted Therapy

14. Liposome Encapsulated Anti-Inflammatory Proteins for targeted Delivery to the Placenta to Treat Fetal Growth Restriction

Padma Murthi and Lynda K. Harris

15. Trophoblast-Targeted Lipsomes for Placenta-Specific Drug Delivery

Part IV Advanced 3D Models

16. Methods for Co-Culture of Primary Human Extravillous Trophoblast Cells and Uterine Natural Killer Cells

Xiaowen Gan, Fen Ning, and Gendie E. Lash

17. Trophoblast Organoids as a Novel Tool to Study Human Placental Development and Function

Sandra Haider, Martin Knöfler, and Paulina A. Latos

18. A Three-Dimensional Trophoblast Invasion Microfluidic Platform for Toxicological Screening

Yong Pu and Almudena Veiga-Lopez

19. Three-Dimensional In Vitro Human Placental Organoids from Mononuclear Villous Trophoblasts or Trophoblast Stem Cells to Understand Trophoblast Dysfunction in Fetal Growth Restriction

Cherry Sun, Joanna L. James, and Padma Murthi

Subject Index List…

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts