Cartilage and Osteoarthritis: Cellular and Molecular Tools

Preface v
Contents of Volume 2 ix
Contributors xi
1 Culture and Phenotyping of Chondrocytes in Primary Culture 1
2 Culture of Chondrocytes in Alginate Beads 15
3 Immortalization of Human Articular Chondrocytes for Generation of Stable,
Differentiated Cell Lines 23
4 Culture of Immortalized Chondrocytes and Their Use As Models of Chondrocyte
Function 37
5 Generation of Pluripotent Stem Cells and Their Differentiation to the
Chondrocytic Phenotype 53
6 Semiquantitative Analysis of Gene Expression in Cultured Chondrocytes by RT-
PCR 69
7 Quantification of mRNA Expression Levels in Articular Chondrocytes With PCR
Technologies 79
8 RNA Extraction From Cartilage 101
9 Gene Expression Analysis in Cartilage by In Situ Hybridization 105
10 Analysis of Differential Gene Expression in Healthy and Osteoarthritic
Cartilage and Isolated Chondrocytes by Microarray Analysis 109
11 High-Efficiency Nonviral Transfection of Primary Chondrocytes 129
12 In Vitro Gene Transfer to Chondrocytes and Synovial Fibroblasts by
Adenoviral Vectors 147
13 Changes of Chondrocyte Metabolism In Vitro: An Approach by Proteomic
Analysis 165
14 Analysis of Chondrocyte Functional Markers and Pericellular Matrix
Components by Flow Cytometry 183
15 A Simple and Reliable Assay of Proteoglycan Synthesis by Cultured
Chondrocytes 209
16 Assays of Proteoglycan and Collagen Degradation in Cultures of Rabbit
Cartilage Explants 219
17 Production of Antibodies Against Degradative Neoepitopes in Aggrecan 237
18 Immunoassays for Collagens in Chondrocyte and Cartilage Explant Cultures 251
19 Detection of Apoptosis in Cartilage In Situ and in Isolated Chondrocytes 275
20 Expression, Activity, and Regulation of MAP Kinases in Cultured Chondrocytes
291
21 Mechanical Loading of Chondrocytes Embedded in 3D Constructs: In Vitro
Methods for Assessment of Morphological and Metabolic Response to Compressive
Strain 307
22 In Vitro Physical Stimulation of Tissue-Engineered and Native Cartilage 325
Index 353