9. Creation of a Bioengineered Ovary: Isolation of Female Germline Stem Cells for the Repopulation of a Decellularized Ovarian Bio-Scaffold
Georgia Pennarossa, Matteo Ghiringhelli, Fulvio Gandolfi, and Tiziana A.L. Brevini
10. A Two-Step Protocol to Erase Human Skin Fibroblasts and Convert Them into Trophoblast-Like Cells
Sharon Arcuri, Fulvio Gandolfi, Edgardo Somigliana, and Tiziana A.L. Brevini
11. 3D-ViaFlow: A Quantitative Viability Assay for Multicellular Spheroids
Joel Mario Vej-Nielsen and Adelina Rogowska-Wrzesinska
12. Method to Disassemble Spheroids into Core and Rim for Downstream Applications Such as Flow Cytometry, Comet Assay, Transcriptomics, Proteomics, and Lipidomics
Helle Sedighi Frandsen, Martina Štampar, Joel Mario Vej-Nielsen, Bojana Žegura, and Adelina Rogowska-Wrzesinska
13. Isolation of Extracellular Vesicles (EVs) Using Size Exclusion-High Performance Liquid Chromatography (SEC-HPLC)
Keerthie Dissanayake, Kasun Godakumara, and Alireza Fazeli
14. Isolation of Extracellular Vesicles (EVs) Using Benchtop Size Exclusion Chromatography (SEC) Columns
Qurat Ul Ain Reshi, Mohammad Mehedi Hasan, Keerthie Dissanayake, and Alireza Fazeli
15. Measurement of the Size and Concentration and Zeta Potential of Extracellular Vesicles Using Nanoparticle Tracking Analyzer
Keerthie Dissanayake, Getnet Midekessa, Freddy Lättekivi, and Alireza Fazeli
16. Isolation, Characterization, and MicroRNAs Analysis of Extracellular Vesicles from Bovine Oviduct and Uterine Fluids
Karina Cañón-Beltrán, Meriem Hamdi, Rosane Mazzarella, Yulia N. Cajas, Claudia L.V. Leal, Alfonso Gutiérrez-Adán, Encina M. González, Juliano C. da Silveira, and Dimitrios Rizos
17. Biomimetic 3D-Bone Tissue Model
Mahmut Parmaksiz, Ayşe Eser Elçin, and Yaşar Murat Elçin
18. Using the Air-Liquid Interface Approach to Foster Apical-Basal Polarization of Mammalian Female Reproductive Tract Epithelia In Vitro
Shuai Chen and Jennifer Schoen
19. Preparation of Biological Scaffolds and Primary Intestinal Epithelial Cells to Efficiently 3D Model the Fish Intestinal Mucosa
Nicole Verdile, Anna Szabo, Rolando Pasquariello, Tiziana A.L. Brevini, Sandra Van Vlierberghe, and Fulvio Gandolfi
20. Use of Porous Polystyrene Scaffolds to Bioengineer Human Epithelial Tissues In Vitro
Lydia Costello, Nicole Darling, Matthew Freer, Steven Bradbury, Claire Mobbs, and Stefan Przyborski
This detailed book collects original protocols aimed at encouraging and stimulating the scientific community to design and produce models for the laboratory that mimic cell guidance conditions as they occur in vivo. The protocols collected describe powerful strategies to exploit chemical cues involved in cell differentiation processes. Special emphasis is given to the use of methods for purification and characterization of exosomes and other secreted vesicles, as well as micro and non-coding RNAs, that have been demonstrated to control the tuning of the in vivo micro and macro environment in order to ensure the optimal soluble environment in vitro. Written for the highly successful Methods in Molecular Biology series, 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.
Authoritative and practical, Next Generation Culture Platforms for Reliable In Vitro Models: Methods and Protocols serves as an ideal guide for researchers working toward developing these vital testing models for preclinical studies.