Chondrocytes Methods and Protocols

This detailed book collects a variety of methods from investigators at the forefront of chondrocyte biology and pathology research that they have tailored for their research projects in order to tackle the unique properties and challenges of the chondrocyte and those of its tissue matrix. Divided into three parts, the volume explores experimental models to study chondrocytes, in vivo assays, as well as methods to characterize the chondrocyte phenotype with high-throughput or discrete assays on cells maintained in vivo, just isolated from their tissue (ex vivo), or cultured in vitro. Written in 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. 

1. Isolation of Chondrocytes from Human Cartilage and Cultures in Monolayer and 3D

            Rodrigo A. Somoza and Jean F. Welter

 

2. Protocol for the Isolation of Intact Chondrons from Healthy and Osteoarthritic Human Articular Cartilage

            Ilona Uzieliene, Jaroslav Denkovskij, Eiva Bernotiene, Ursule Kalvaityte, Raminta Vaiciuleviciute, Yolande F.M. Ramos, and Ali Mobasheri

 

3. Cartilage-Specific Cre Recombinase Transgenes/Alleles in the Mouse

            Ioannis Kanakis, Mohammad Alhashmi, Ke Liu, Craig Keenan, Lorenzo Ramos Mucci, Blandine Poulet, and George Bou-Gharios

 

4. Isolation of Mouse Growth Plate and Articular Chondrocytes for Primary Cultures

            Abdul Haseeb and Véronique Lefebvre

 

5. Fetal Growth Plate Cartilage: Histological and Immunohistochemical Techniques

            Zachary Tata, Christophe Merceron, and Ernestina Schipani

 

6. Preparation of Adult Mouse Skeletal Tissue Sections for RNA In Situ Hybridization

            Charles de Charleroy, Abdul Haseeb, and Véronique Lefebvre

 

7. MicroRNA In Situ Hybridization in Paraffin-Embedded Human Articular Cartilage and Mouse Knee Joints

            Helal Endisha and Mohit Kapoor

 

8. Laser Capture Microdissection of Mouse Growth Plate Cartilage

            Bijal Kikani and Julian C. Lui

 

9. RNA Isolation from Articular Cartilage Tissue

            Hongjun Zheng and Audrey McAlinden

 

10. Small Nucleolar RNA Expression Profiling in Cartilage

            Mandy J. Peffers, Andy Cremers, and Tim J.M. Welting

 

11. MicroRNA Expression Profiling, Target Identification, and Validation in Chondrocytes

            Tomoki Chiba, Ryota Kurimoto, Takahide Matsushima, Yoshiaki Ito, Ryo Nakamichi, Martin Lotz, and Hiroshi Asahara

 

12. ChIP-Seq Assays from Mammalian Cartilage and Chondrocytes

            Akira Yamakawa, Hironori Hojo, and Shinsuke Ohba

 

13. MicroRNAs and Regulation of Autophagy in Chondrocytes

            Yolande F.M. Ramos and Ali Mobasheri

 

14. Mitochondrial Biogenesis, Activity, and DNA Isolation in Chondrocytes

            Yun Wang, Liang-Yu Chen, and Ru Liu-Bryan

 

15. Assessing Chondrocyte Status by Immunofluorescence-Mediated Localization of Parkin Relative to Mitochondria

            Mohammad Y. Ansari and Tariq M. Haqqi