Main description:

Skeletal Development and Repair: Methods and Protocols is a compilation of a variety of skeletal research protocols utilizing the laboratory mouse as the platform for surgical manipulation and/or transplantation as well as the source of tissues and cells for in vitro culture and analyses. Chapters are written by experts in the field and cover topics including surgical, transplantation and organ culture methods that permit analyses of skeletal tissues undergoing repair in vivo and permits analyses of cellular interactions ex vivo, histological and molecular techniques developed to study gene and protein expression in whole embryos, skeletal tissues and tissue sections and in vitro primary cell culture protocols designed to assay gene function in specific cell populations. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Skeletal Development and Repair: Methods and Protocols is a comprehensive laboratory manual for all levels of basic research scientists working in the broad fields of skeletal development and skeletal repair research.

Contents:

Part I. Understanding Mouse Skeletal Development and Repair

1. Overview of Skeletal Development

Tatsuya Kobayashi and Henry M. Kronenberg

2. Overview of Skeletal Repair (Fracture Healing and Its Assessment)

Elise F. Morgan, Anthony De Giacomo, and Louis C. Gerstenfeld

Part II. Skeletal Repair, Transplantations, and Organ Cultures

3. Generation of Closed Transverse Fractures in Small Animals

Anthony De Giacomo, Elise F. Morgan, and Louis C. Gerstenfeld

4. The Murine Femoral Bone Graft Model and a Semi-Automated Histomorphometric Analysis Tool

Robinder S. Dhillon, Longze Zhang, Edward M. Schwarz, Brendan F. Boyce, and Chao Xie

5. Surgical Induction of Posttraumatic Osteoarthirits

Daisuke Hamada, Erik R. Sampson, Robert D. Maynard, and Michael J. Zuscik

6. A Mouse Model of Flexor Tendon Repair

Sys Hasslund, Regis J. O'Keefe, and Hani A. Awad

7. Cartilage Explant Cultures

Manuela Wuelling and Andrew Vortkamp

8. Renal Capsule Transplantations to Assay Skeletal Angiogenesis

Rana Abou-Khalil and Celine Colnot

Part III. Histological and Molecular Analyses on Whole Tissues and Tissue Sections

9. Whole Mount Skeletal Staining

Diana Rigueur and Karen M. Lyons

10. Tips and Techniques for Processing and Sectioning Undecalcified Murine Bone Specimens

Thomas B. Bemenderfer, Jonathan S. Harris, Keith W. Condon, and Melissa A. Kacena

11. Preparation of Thin Sections from Non-fixed and Undecalcified Hard Tissues Using Kawamoto's Film Method (2012)

Tadafumi Kawamoto and Komei Kawamoto

12. Demineralized Murine Skeletal Histology

Sarah A. Mack, Kathleen M. Maltby, and Matthew J. Hilton

13. Beta-Galactosidase Staining in the Skeleton

X.L. Han and Jian Q. Feng

14. Whole-Mount In Situ Hybridization on Murine Skeletogenic Tissues

Timothy Rutkowsky, Deepika Sharma, and Matthew J. Hilton

15. Non-Radioactive In Situ Hybridization on Skeletal Tissue Sections

Yulia Shwartz and Elazar Zelzer

16. Radioactive In Situ Hybridization to Detect Gene Expression in Skeletal Tissue Sections

Kyu Sang Joeng, Jenna Regan, and Fanxin Long

17. Proliferation Assays (BrdU and EdU) on Skeletal Tissue Sections

Timothy J. Mead and Veronique Lefebvre

18. TUNEL Assay on Skeletal Tissue Sections to Detect Apoptosis

Laura Mangiavini and Ernestina Schipani

Part IV. Primary Cell Isolations, Cultures, and Assays

19. Analysis of Chondrogenesis Using Micromass Cultures of Limb Mesenchyme

T. Michael Underhill, Helen J. Dranse, and Lisa M. Hoffman

20. Isolation and Culture of Murine Primary Chondrocytes

Anthony J. Mirando, Yufeng Dong, Jinsil Kim, and Matthew J. Hilton

21. Bone Marrow Stromal Cell Assays - In Vitro and In Vivo

Pamela Gehron Rober, Sergei A. Kuznetsov, Mara Riminucci, and Paolo Bianco

22. Isolation and Culture of Neonatal Mouse Calvarial Osteoblasts

Jennifer H. Jonason and Regis J. O'Keefe

23. RANKL-Based Osteoclastogenic Assays from Murine Bone Marrow Cells

Lianping Xing and Brendan F. Boyce

24. Hematopoietic Stem Cell Cultures and Assays

Benjamin J. Frisch and Laura M. Calvi