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The AdLMP-1 transfection in two different cells; AF cells, chondrocytes as potential cell therapy candidates for disc degeneration.

Authors
 Kuh SU  ;  Zhu Y  ;  Li J  ;  Tsai KJ  ;  Fei Q  ;  Hutton WC  ;  Yoon ST 
Citation
 ACTA NEUROCHIRURGICA, Vol.150(8) : 803-810, 2008 
Journal Title
 ACTA NEUROCHIRURGICA 
ISSN
 0001-6268 
Issue Date
2008
MeSH
Adaptor Proteins, Signal Transducing ; Aggrecans/genetics ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Protein 7 ; Bone Morphogenetic Proteins/metabolism ; Cell Transplantation* ; Chondrocytes/metabolism* ; Chondrocytes/pathology ; Chondrocytes/transplantation* ; Collagen Type I/genetics ; Collagen Type II/genetics ; Cytoskeletal Proteins ; Gene Expression Regulation/genetics ; Genetic Therapy/methods* ; Glycosaminoglycans/metabolism ; Humans ; Intervertebral Disc/metabolism* ; Intervertebral Disc/pathology ; Intracellular Signaling Peptides and Proteins/genetics* ; LIM Domain Proteins ; Polymerase Chain Reaction ; RNA, Messenger/genetics ; Spinal Diseases/therapy* ; Transfection/methods* ; Transforming Growth Factor beta/metabolism
Keywords
Proteoglycan ; Anulus fibrosus cell ; Chondrocyte ; AdLMP-1
Abstract
BACKGROUND: LMP-1 is known to increase proteoglycan production through the upregulating the BMPs and it is also known that BMP-2 acts on anulus fibrosus cells and chondrocytes to increase proteoglycan production. METHOD: We carried out an experiment, the effect of AdLMP-1 transfection on AF cells and chondrocytes in the production of sulfated-glycosaminoglycans, mRNA expression (aggrecan, type I, II collagen, LMP-1, BMP-2, and BMP-7), and immunofluorescence staining. AF cells and chondrocytes were grown in monolayer and treated for 6 days with AdLMP1-green fluorescence protein (GFP) (10, 20, and 30 multiplicity of infection [MOI]). After 6 days, the sGAG content in the media was quantified using 1,9-dimethylmethylene blue staining. The mRNA expression was measured with real-time PCR after 20 MOI infection of AdLMP1-GFP. The each cells treated with 20 MOI infection of AdGFP was used as a control group for the mRNA expression. The each cell group was immunofluorescence stained with each antibodies in the chamber slide at 3 x 10(4) cells/chamber. FINDINGS: 1) The sGAG production was maximum in 20 MOI AdLMP1-GFP infection on the AdLMP-1 treatment for both of AF cells and chondrocytes. 2) The mRNA expression of aggrecan, type I collagen, type II collagen, LMP-1, BMP-2, and BMP-7 is increased in both AF cells and chondrocytes in 20 MOI AdLMP1-GFP infection. 3) On the immunofluorescence staining results, the positive immunofluorescence stained cell numbers are increased after 20 MOI AdLMP1-GFP infection concordant with upregulation of mRNA expression. CONCLUSIONS: The AdLMP-1 treatments in AF cells and chondrocytes may be useful for cell transplantation therapy in disc degeneration
Full Text
http://link.springer.com/article/10.1007/s00701-008-1617-7
DOI
10.1007/s00701-008-1617-7
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Neurosurgery (신경외과학교실) > 1. Journal Papers
Yonsei Authors
Kuh, Sung Uk(구성욱) ORCID logo https://orcid.org/0000-0003-2566-3209
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/106871
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