Successful in vitro studies of disc cells require interaction of the c
ell with a compatible microenvironment which favors expression of the
disc cell phenotype. The objective of this study was to characterize c
ells grown by standardized methods of isolation and passage, and cultu
re cells from the human annulus in three-dimensional culture. Cells fr
om the annulus of 11 individuals were cultured in alginate or agarose
for ten days, and extracellular matrix components were evaluated with
immunohistochemistry and quantitative analysis of the percent of colon
ies producing Type I or II collagen, 4-sulfated chondroitin sulfate or
keratan sulfate. Results show production of these four extracellular
matrix products through multiple passages and support the phenotypic s
tability of disc cells in three-dimensional culture.