Progenitor cells : methods and protocols

This volume explores the diversity in progenitor cell biology methods and uniquely describes techniques for isolating, generating, and characterizing progenitor cells from either tissue or embryonic stem cells. The chapters in this book cover topics such as isolating progenitor cells from sources including adipose tissue, blood, bone marrow, ear, gut, heart, pancreatic islets, and Whirton’s jelly; deriving neural, pancreatic, and urothelial precursors from the embryonic stem cells; and culturing isolated islets in vitro to generate progenitor cells via epithelial-to-mesenchymal transition. This book also delves into digital droplet PCR; flow cytometry and cell sorting; mitochondrial assays; calcium ratiometric signaling; and 2D gel electrophoresis. 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, __Progenitor Cells: Methods and Protocols__ is a valuable resource for scientists and researchers interested in learning more about this evolving and diverse field. Front Matter ....Pages i-xiv Isolation and Molecular Characterization of Progenitor Cells from Human Umbilical Cord (Umesh Goyal, Ankita Sen, Malancha Ta)....Pages 1-13 Isolation and Characterization of Extracellular Vesicles from Mesenchymal Stromal Cells (Sally Yunsun Kim, Thanh Huyen Phan, Christina Limantoro, Bill Kalionis, Wojciech Chrzanowski)....Pages 15-23 Assessment of β-Cell Replication in Isolated Rat Islets of Langerhans (Louise T. Dalgaard)....Pages 25-35 Droplet Digital PCR for Measuring Absolute Copies of Gene Transcripts in Human Islet-Derived Progenitor Cells (Cody-Lee Maynard, Wilson K. M. Wong, Anandwardhan A. Hardikar, Mugdha V. Joglekar)....Pages 37-48 Isolation and In Vitro Culture of Human Gut Progenitor Cells (Jessica Bruce, Gerard E. Kaiko, Simon Keely)....Pages 49-62 Isolation and Characterization of Colony-Forming Progenitor Cells from Adult Pancreas (Janine C. Quijano, Jacob R. Tremblay, Jeffrey Rawson, Hsun Teresa Ku)....Pages 63-80 A Novel Gene Delivery Approach Using Metal Organic Frameworks in Human Islet-Derived Progenitor Cells (Arpita Poddar, Mugdha V. Joglekar, Anandwardhan A. Hardikar, Ravi Shukla)....Pages 81-91 Encapsulation and Transplantation of Pancreatic Progenitor Cells (Luke Carroll, Auvro R. Mridha, Bernard E. Tuch)....Pages 93-102 Differentiation of Urothelium from Mouse Embryonic Stem Cells in Chemically Defined Conditions (Badwi B. Boumelhem, Stuart T. Fraser, Stephen J. Assinder)....Pages 103-115 Isolation and Characterization of Progenitor Cells from Human Adipose Tissue (Nitya Shree, Ramesh Bhonde)....Pages 117-123 Identification and Analysis of Mouse Erythroid Progenitor Cells (Chanukya K. Colonne, Jia Hao Yeo, Campbell V. McKenzie, Stuart T. Fraser)....Pages 125-145 Single-Cell Assays Using Hematopoietic Stem and Progenitor Cells (Ashwini S. Hinge, Marie-Dominique Filippi)....Pages 147-160 Isolation and Characterization of Cardiac Progenitor Cells (Parul Dixit, Rajesh G. Katare)....Pages 161-173 Mitochondrial Assays Using Cardiac Stem Cells (Ayeshah A. Rosdah, Lea M. D. Delbridge, Shiang Y. Lim)....Pages 175-183 Two-Dimensional Electrophoresis and Mass Spectrometry for Protein Identification (Amaresh K. Ranjan, Anil Gulati)....Pages 185-195 High-Efficiency Lentiviral Gene Modification of Primary Murine Bone-Marrow Mesenchymal Stem Cells (Dario Gerace, Binhai Ren, Rosetta Martiniello-Wilks, Ann M. Simpson)....Pages 197-214 Bone Marrow-Derived Progenitor Cells Mediate Immune Cell Regulation (Kisha N. Sivanathan, Patrick T. Coates)....Pages 215-234 Flow Cytometry and Cell Sorting Using Hematopoietic Progenitor Cells (Sangeetha Vadakke-Madathil, Lalita S. Limaye, Vaijayanti P. Kale, Hina W. Chaudhry)....Pages 235-246 Isolation of Epidermal Progenitor Cells from Rat Tympanic Membrane (Lawrence J. Liew, Allen Y. Wang, Rodney J. Dilley)....Pages 247-255 Measurement of Store-Operated Calcium Entry in Human Neural Cells: From Precursors to Differentiated Neurons (Renjitha Gopurappilly, Bipan Kumar Deb, Pragnya Chakraborty, Gaiti Hasan)....Pages 257-271 Embryoid Body Differentiation of Mouse Embryonic Stem Cells into Neurectoderm and Neural Progenitors (Rachel A. Shparberg, Hannah J. Glover, Michael B. Morris)....Pages 273-285 Back Matter ....Pages 287-289 This volume explores the diversity in progenitor cell biology methods and uniquely describes techniques for isolating, generating, and characterizing progenitor cells from either tissue or embryonic stem cells. The chapters in this book cover topics such as isolating progenitor cells from sources including adipose tissue, blood, bone marrow, ear, gut, heart, pancreatic islets, and Whirton’s jelly; deriving neural, pancreatic, and urothelial precursors from the embryonic stem cells; and culturing isolated islets in vitro to generate progenitor cells via epithelial-to-mesenchymal transition. This book also delves into digital droplet PCR; flow cytometry and cell sorting; mitochondrial assays; calcium ratiometric signaling; and 2D gel electrophoresis. 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, Progenitor Cells: Methods and Protocols is a valuable resource for scientists and researchers interested in learning more about this evolving and diverse field.