Differing in vitro biology of equine, ovine, porcine and human articular chondrocytes derived from the knee joint: an immunomorphological study.
For lack of sufficient human cartilage donors, chondrocytes isolated from various animal species are used for cartilage tissue engineering. The present study was undertaken to compare key features of cultured large animal and human articular chondrocytes of the knee joint. Primary chondrocytes were isolated from human, porcine, ovine and equine full thickness knee joint cartilage and investigated flow cytometrically for their proliferation rate. Synthesis of extracellular matrix proteins collagen type II, cartilage proteoglycans, collagen type I, fibronectin and cytoskeletal organization were studied in freshly isolated or passaged chondrocytes using immunohistochemistry and western blotting. Chondrocytes morphology, proliferation, extracellular matrix synthesis and cytoskeleton assembly differed substantially between these species. Proliferation was higher in animal derived compared with human chondrocytes. All chondrocytes expressed a cartilage-specific extracellular matrix. However, after monolayer expansion, cartilage proteoglycan expression was barely detectable in equine chondrocytes whereby fibronectin and collagen type I deposition increased compared with porcine and human chondrocytes. Animal-derived chondrocytes developed more F-actin fibers during culturing than human chondrocytes. With respect to proliferation and extracellular matrix synthesis, human chondrocytes shared more similarity with porcine than with ovine or equine chondrocytes. These interspecies differences in chondrocytes in vitro biology should be considered when using animal models.
Department of Trauma and Reconstructive Surgery, Charité-Campus Benjamin Franklin, FEM, Krahmerstrasse 6-10, 12207, Berlin, Germany. gundula.schulze-tanzil@charite.de
G
G
Schulze-Tanzil
R D
RD
Müller
B
B
Kohl
N
N
Schneider
W
W
Ertel
K
K
Ipaktchi
H
H
Hünigen
O
O
Gemeinhardt
R
R
Stark
T
T
John
D000199:Actins / Q000032:analysis; D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D003599:Cytoskeleton; D016326:Extracellular Matrix Proteins / Q000032:analysis; D006736:Horses; D006801:Humans; D007719:Knee Joint; D012756:Sheep; D013045:Species Specificity; D013552:Swine
D000199:Actins; D016326:Extracellular Matrix Proteins