Versican G3 domain enhances cellular adhesion and proliferation of bovine intervertebral disc cells cultured in vitro.
The functional role of versican in influencing intervertebral disc cell adhesion and proliferation was analyzed in bovine intervertebral disc. We have previously demonstrated the C-terminal globular G3 (or selectin-like) domain of versican to influence mesenchymal chondrogenesis and fibroblast proliferation in vitro. For this study, a versican G3 expression construct was generated to examine the role of the G3 domain of versican. Nucleus pulposus and annulus fibrosus cells were isolated from adult bovine caudal discs using sequential enzymatic digestion and versican expression characterized by RT-PCR. In cell proliferation assays, we observed that there was greater cellular proliferation in the presence of versican G3 for both disc cell types. The higher proliferation rate of annulus fibrosus cells when compared to nucleus pulposus cells seeded in monolayer supports heterogeneity of intervertebral disc cell populations. The presence of versican G3 construct enhanced the adhesion of isolated nucleus pulposus and annulus fibrosus cells approximately 4 to 6 fold, respectively. Cellular adhesion was greater in the presence of versican G3 in a dose dependent manner. G3 product was purified using affinity columns, and the purified G3 also enhanced cell adhesion.
Yang
Bing L
BL
Sunnybrook and Women's College Health Sciences Centre, and University of Toronto, Toronto, Ontario, Canada.
Yang
Burton B
BB
Erwin
Mark
M
Ang
Lee Cyn
LC
Finkelstein
Joel
J
Yee
Albert J M
AJ
D000818:Animals; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D011508:Chondroitin Sulfate Proteoglycans / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression; D066298:In Vitro Techniques; D007403:Intervertebral Disc / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D037181:Lectins, C-Type; D017434:Protein Structure, Tertiary; D011509:Proteoglycans / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053675:Versicans
D011508:Chondroitin Sulfate Proteoglycans; D017077:Culture Media, Conditioned; D037181:Lectins, C-Type; D011509:Proteoglycans; D012333:RNA, Messenger; D053675:Versicans