Studies of human intervertebral disc cell function in a constrained in vitro tissue culture system.
METHODS
This is a laboratory-based study examining a novel in vitro culture system for intervertebral disc tissue.


OBJECTIVE
Address the hypothesis that "the novel culture system will preserve intervertebral disc tissue matrix and cell function and prevent cellular apoptosis for periods up to 21 days."


BACKGROUND
Studies of cell function in human intervertebral disc tissue are scarce. In vivo study of human intervertebral disc cells remains impracticable; in situ molecular biology in histologic sections lacks a dynamic dimension; and as for in vitro studies, cell culture often lacks physiologic relevance and explant cultures are subject to loss of tissue integrity and altered cell behavior. There is a biologic and therapeutic need for a satisfactory explant culture system for studying human intervertebral disc tissue in a controlled environment.


METHODS
Samples of human intervertebral disc tissue, obtained at surgery, were examined for a number of tissue and cell parameters immediately after excision (controls) and following culture of tissue samples either in a plastic ring or unconstrained in tissue culture medium for up to 3 weeks. Data were compared between cultured tissue and controls.


RESULTS
By comparison with control tissue, unconstrained explants swelled, tissue structure was disturbed, and there were profound changes in cell function. By contrast, tissue cultured in plastic rings maintained tissue structure, and after 3 weeks, the cellular parameters were the same as in controls.


CONCLUSIONS
This is the first reported system to preserve cell function of human discal explants for long periods in tissue culture. It will be a useful tool for a wide range of investigations of intervertebral disc biology that have not hitherto been possible.
Le Maitre
Christine Lyn
CL
Laboratory Medicine Academic Group, University of Manchester, Manchester, United Kingdom. christine.lemaitre@man.ac.uk
Hoyland
Judith Alison
JA
Freemont
Anthony J
AJ
D000328:Adult; D053674:Aggrecans; D000423:Alcian Blue; D017209:Apoptosis; D003094:Collagen / Q000378:metabolism; D004396:Coloring Agents; D046508:Culture Techniques / Q000379:methods*; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism; D016326:Extracellular Matrix Proteins; D006609:High Mobility Group Proteins / Q000235:genetics / Q000378:metabolism; D006651:Histocytochemistry; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007403:Intervertebral Disc / Q000033:anatomy & histology / Q000166:cytology* / Q000378:metabolism; D037181:Lectins, C-Type; D008875:Middle Aged; D011509:Proteoglycans / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D055758:SOX9 Transcription Factor; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D053674:Aggrecans; D004396:Coloring Agents; D016326:Extracellular Matrix Proteins; D006609:High Mobility Group Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D012333:RNA, Messenger; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D014157:Transcription Factors; D003094:Collagen; D000423:Alcian Blue