Proliferation of canine intervertebral disk chondrocytes in three-dimensional alginate microsphere culture.
Proliferation of chondrocytes from nucleus pulposus (NP) and anulus fibrosus (AF) was confirmed in three-dimensional culture using alginate microspheres. Cells isolated from NP and AF were incorporated in microspheres and cultured for 14 days. Round mononuclear cells of 20-25 microm in diameter proliferated and formed aggregates. At day 14, alcian blue positive matrix surrounded the proliferating cells. The cells had cytoplasmic vacuoles stained positively by toluidine blue. On electron microscopy, the cells contained proteoglycan vacuoles and lipid droplets in the cytoplasm and synthesized collagen fibrils and electron dense granules surrounding the cell. These features of the cells were characteristic for chondrocytes. This culture system should be useful to further investigate metabolic activities of intervertebral disk chondrocytes.
Masuda
Kenichi
K
Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana 61802, USA.
Miyabayashi
Takayoshi
T
Meachum
Sharon H
SH
Eurell
Thomas E
TE
D000423:Alcian Blue / Q000737:chemistry; D000464:Alginates / Q000737:chemistry*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods / Q000662:veterinary; D002455:Cell Division / Q000502:physiology; D019902:Chondrocytes / Q000166:cytology* / Q000648:ultrastructure; D004283:Dog Diseases / Q000473:pathology; D004285:Dogs / Q000502:physiology*; D005260:Female; D007403:Intervertebral Disc / Q000166:cytology*; D008297:Male; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D008863:Microspheres
D000464:Alginates; D000423:Alcian Blue