Cell-Cycle Synchronization Methods and Protocols

This volume covers a broad range of cell types including cultured cell lines, primary cells, and various unicellular organisms such as fission yeast, budding yeast, parasite Leishmania amazonensis, and parasite Trypanosoma brucei. The chapters in this book are organized into four parts. Part One looks at a general overview of cell cycle control and synchronization. Part Two discusses techniques to synchronize mammalian cells to various cell cycle phases including mitotic sub-phases. Part Three covers synchronization of unicellular organisms and Part Four analyzes cell cycle progression. 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, Cell-Cycle Synchronization: Methods and Protocols is a valuable resource for both novice and expert scientists in this developing field.

Part I   Introduction and Review

1.     Cell Cycle Progression and Synchronization: An Overview

Zhixiang Wang

2.     The Trypanosomatids Cell Cycle: A Brief Report
Arthur de Oliveira Passos, Luiz H. C. Assis, Yete G. Ferri, Vitor L. da Silva, Marcelo S. da Silva, Maria Isabel N. Cano

3.     Cell Cycle-Related Clinical Applications
Md Gulam Musawwir Khan and Yi Wang

4.     Flow Cytometry and Cell Cycle Analysis: An Overview

            Aja M. Rieger

Part II Methods for Cell Cycle Synchronization of Mammalian Cells

5.     Synchronization of Cultured Cells to G1, S, G2, and M Phases by Double Thymidine Block

Richard C. Wang and Zhixiang Wang

Joanne D. Hadfield1, Sargun Sokhi, and Gordon K. Chan

7.     Synchronization of HeLa Cells to Various Interphases Including G1, S, and G2 Phases
Ping Wee, Richard C. Wang, and Zhixiang Wang

8.     Synchronization of HeLa Cells to Mitotic Subphases

Ping Wee, Richard C. Wang, and Zhixiang Wang

9.     Cell Cycle Synchronization of Primary and Cultured Articular Chondrocytes

Part III Methods for Cell Cycle Synchronization of Unicellular Organisms

10.  Synchronization of Leishmania amazonensis Cell Cycle using Hydroxyurea
Beatriz C. D. Oliveira, Luiz H. C. Assis, Mark E. Shiburah, Stephany C. Paiva, Veronica S. Fontes, Leilane S. de Oliveira, Vitor L. da Silva, Marcelo S. da Silva, Maria Isabel N. Cano

11.  Synchronization of Trypanosoma brucei by Counter-Flow Centrifugal Elutriation

Corinna Benz and Michael D. Urbaniak

12.  Synchronization of Saccharomyces cerevisiae Cells for Analysis of Progression through the Cell Cycle

Brianna L. Greenwood and David T. Stuart

13.  Cell Cycle Synchrony Methods for Fission Yeast, Schizosaccharomyces pombe

Zohreh Kianfard, Kyle Cheung, and Sarah A. Sabatinos

Part IV Methods to Assess Cell Cycle Progression

14.  Analysis of Cell Cycle by Flow Cytometry

Hamid Maadi, Mohammad Hasan Soheilifar, and Zhixiang Wang

15.  Analysis of Cell Proliferation by Three-Dimensional Culture

Xiaoyun Tang and David N. Brindley

16.  BrdU Incorporation Assay to Analyze the Entry into S Phase

Jihang Yu, Zhixiang Wang, and Yi Wang

17.  Assessment of Growth Plate Chondrocytes Proliferative Activity in Embryonic Endochondral Ossification via Ki67 Immunofluorescence

Asra Almubarak and Fred B. Berry

18.  Detecting Cell Cycle Stage and Progression in Fission Yeast, Schizosaccharomyces pombe

Zohreh Kianfard, Kyle Cheung, Daniel Rappaport, Sirasie Magalage, and Sarah A. Sabatinos