Differential expression of galectin-1 and its interactions with cells and laminins in the intervertebral disc.
Galectin-1 (Gal-1), an endogenous β-galactoside-binding protein, binds to laminins, which are highly expressed in the nucleus pulposus (NP) of the intervertebral disc (IVD). The objective of this study is to evaluate the expression of Gal-1 protein in IVD tissues during aging and the effect of Gal-1 on IVD cell adhesion to laminins. Tissues from rat, porcine, and human (scoliosis or disc degeneration) IVDs were used to evaluate Gal-1 expression via immunostaining, RT-PCR, and Western blot analysis. Attachment of isolated IVD cells (porcine and human) on select laminin isoforms (LM-111 and LM-511) was compared with/without pre-incubation with exogenous Gal-1. A biotinylated Gal-1(B-Gal-1) was used to evaluate for binding to IVD cells and to select for IVD cells by magnetic activated cell sorting (MACS). NP cells expressed high levels of Gal-1 protein as compared to anulus fibrosus (AF) cells in immature tissues, while exogenous Gal-1 increased both NP and AF cell attachment to laminins and exhibited a similar binding to both cell types in vitro. With aging, Gal-1 levels in NP tissue appeared to decrease. In addition, incubation with B-Gal-1 was able to promote the retention of more than 50% of IVD cells via MACS. Our results provide new findings for the presence and functional role of Gal-1 within IVDs. Similar staining patterns for Gal-1 and LM-511 in IVD tissue suggest that Gal-1 may serve as an adhesion molecule to interact with both cells and laminins. This MACS protocol may be useful for selecting pure IVD cells from mixed cells of pathological tissue.
Department of Biomedical Engineering, Duke University, Durham, North Carolina 27708, USA.
Liufang
L
Jing
Stephen
S
So
Shaun W
SW
Lim
William J
WJ
Richardson
Robert D
RD
Fitch
Lori A
LA
Setton
Jun
J
Chen
D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D019921:Biotinylation; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D002648:Child; D005109:Extracellular Matrix / Q000378:metabolism; D037483:Galectin 1 / Q000096:biosynthesis*; D005786:Gene Expression Regulation; D006801:Humans; D007403:Intervertebral Disc / Q000254:growth & development / Q000378:metabolism*; D007797:Laminin / Q000378:metabolism*; D008875:Middle Aged; D008954:Models, Biological; D013552:Swine
D015815:Cell Adhesion Molecules; D037483:Galectin 1; D007797:Laminin