Cells from different regions of the intervertebral disc: effect of culture system on matrix expression and cell phenotype.
METHODS
This study examined how the culture system and region of cellular origin affect disc cell morphology and extracellular matrix production.


OBJECTIVE
To determine the role of the cell populations in the different regions of the adult intervertebral disc in maintaining gradients in composition across the disc.


BACKGROUND
It is not known whether the steep profiles in composition across the intervertebral disc are maintained by distinct cell populations or whether differences in cell metabolism are determined by changes in the physical environment across the disc. Very little information exists on the matrix produced by cells from the mature, non-notochordal nucleus pulposus.


METHODS
Cells were extracted from articular cartilage, nucleus pulposus, and the inner and outer anulus fibrosus of caudal discs from 18- to 24-month-old steers cultured in alginate or collagen gels or in monolayer. The effect of culture system and cell origin on cell morphology and matrix synthesis was measured using 35S-sulphate labeling and indirect immunolocalization.


RESULTS
Distinct morphologic differences between cells from different regions cultured in monolayer were retained through two passages. The rate of sulfate incorporation varied with cell type. Immediately after isolation, it was two- to threefold greater for nucleus cells than for cells from the disc inner anulus or articular cartilage. The rate was lowest for outer anulus cells. It also varied with culture system. For all cell types, the incorporation rate was highest in alginate and lowest in monolayer. Immunolocalization showed that nucleus cells stained strongly for all proteoglycan epitopes, whereas outer anulus cells stained least and in monolayer produced little proteoglycan.


CONCLUSIONS
The disc has at least three distinct cell populations, which differ in morphology and in amount and type of matrix they produce. Cells from mature nucleus pulposus produced sulfated glycosaminoglycans at a high rate in contrast to reported results for notochordal nucleus cells. Alginate, although an appropriate culture system for inner anulus and nucleus cells, may not be a suitable medium for outer anulus cells.
Horner
Heather A
HA
Physiology Laboratory, Oxford University, Oxford, and the; Centre for Spinal Studies, RJAH Orthopaedic Hospital, Oswestry, Shropshire, United Kingdom.
Roberts
Sally
S
Bielby
Robert C
RC
Menage
Janis
J
Evans
Helen
H
Urban
Jill P G
JP
D000818:Animals; D002358:Cartilage, Articular / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D002417:Cattle; D048429:Cell Size; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism; D007150:Immunohistochemistry; D007403:Intervertebral Disc / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D008297:Male; D011509:Proteoglycans / Q000032:analysis; D013431:Sulfates / Q000378:metabolism
D011509:Proteoglycans; D013431:Sulfates