Primary culture and phenotyping of murine chondrocytes

Nature Protocols
Marjolaine GossetClaire Jacques

Abstract

The culture of chondrocytes is one of the most powerful tools for exploring the intracellular and molecular features of chondrocyte differentiation and activation. However, chondrocytes tend to dedifferentiate into fibroblasts when they are subcultured, which is a major problem. This protocol, involving primary cultures to limit dedifferentiation, describes two different methods for culturing chondrocytes of different anatomical origins (articular and costal chondrocytes, both of which represent hyaline cartilage) from mice. Mice are of particular interest for cellular and molecular studies, as many tools suitable for use in mice are available. In addition, rapid development of transgenic and gene-targeted mice provides powerful instruments for biological studies. The protocol can be divided into four stages: isolation of cartilage (15 min per animal), isolation of chondrocytes (2 h extended overnight), seeding of chondrocytes (1 h 30 min) and growth in culture (6 d). To obtain confluency of chondrocytes using this protocol takes 7 d. Methods for phenotyping chondrocytes are also provided.

References

Apr 1, 1992·Matrix : Collagen and Related Research·H J HäuselmannE J Thonar
Apr 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·F Mallein-Gerin, B R Olsen
Aug 1, 1994·Matrix Biology : Journal of the International Society for Matrix Biology·V LefebvreB De Crombrugghe
Dec 1, 1994·The Journal of Clinical Investigation·M B GoldringJ F Apperley
Feb 1, 2000·The Journal of Biological Chemistry·S MurakamiB de Crombrugghe
Feb 3, 2000·Proceedings of the National Academy of Sciences of the United States of America·S MurakamiB de Crombrugghe
Sep 12, 2000·Matrix Biology : Journal of the International Society for Matrix Biology·B de CrombruggheW Huang
Oct 3, 2000·Arthritis and Rheumatism·M B Goldring
Feb 25, 2005·Osteoarthritis and Cartilage·Colette SalvatSylvie Thirion

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Citations

Jun 1, 2013·Inflammation Research : Official Journal of the European Histamine Research Society ... [et Al.]·Fazal-Ur-Rehman BhattiSheikh Riazuddin
Apr 2, 2013·Journal of Molecular Histology·Weiliang ZhongXifu Zheng
Jul 9, 2009·Molecular Therapy : the Journal of the American Society of Gene Therapy·Fan YangKenneth M C Cheung
Jan 16, 2013·Proceedings of the National Academy of Sciences of the United States of America·Yoko HosakaHiroshi Kawaguchi
Nov 2, 2012·Proceedings of the National Academy of Sciences of the United States of America·Brian O DiekmanFarshid Guilak
Jun 5, 2013·Molecular and Cellular Biology·Katherine C HallCarl P Blobel
Jan 20, 2012·Annals of the Rheumatic Diseases·Odile GabayMona Dvir-Ginzberg
Feb 5, 2011·The Journal of Clinical Investigation·Kunihiko HiramatsuNoriyuki Tsumaki
Apr 4, 2013·The Journal of Clinical Investigation·Kapil V RamachandranGeoffrey S Pitt
May 31, 2014·PloS One·Glyn D PalmerSteven B Abramson
Nov 3, 2010·The Tohoku Journal of Experimental Medicine·Jie ChengYunong Wu
Feb 1, 2014·Arthritis Research & Therapy·Marie-Charlotte LaiguillonJérémie Sellam
Feb 8, 2014·Canadian Journal of Physiology and Pharmacology·Zhiqiang YeDaozhang Cai
Sep 19, 2014·Nature·Akihiro YamashitaNoriyuki Tsumaki
Dec 15, 2015·Journal of Cellular Physiology·Hyunjun ShinJennifer H Shin
Mar 10, 2016·Materials Science & Engineering. C, Materials for Biological Applications·A ThiemR Zehbe
Mar 3, 2015·Proceedings of the National Academy of Sciences of the United States of America·Makoto HirataBenjamin A Alman
Mar 27, 2012·Molecular Immunology·Hong-Hsing LiuGary Peltz

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