Linked Life Data

11561005

Source:http://linkedlifedata.com/resource/pubmed/id/11561005

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
10
pubmed:dateCreated
2001-9-18
pubmed:abstractText
Culture of articular chondrocytes in alginate beads offers several advantages over culture in monolayer; cells retain their phenotype for 8 months or longer. Earlier studies of chondrocytes cultured in alginate concentrated on collagen and proteoglycan synthesis. However, gene expression by in situ hybridization (ISH) has not been investigated. The purposes of the present study on human chondrocytes were (a) to modify the ISH procedure for the alginate beads to examine the mRNA expression of alpha1 (II) procollagen, aggrecan, and two matrix metalloproteinases (MMP-3 and MMP-8) thought to be involved in cartilage matrix degradation, and (b) to compare expression in cultured chondrocytes with that in chondrocytes of intact human cartilage. The modifications made for ISH include the presence of CaCl2 and BaCl2 in the fixation and washing steps and exclusion of cetyl pyridinium chloride. By ISH we show that aggrecan, MMP-3, and MMP-8 are continuously expressed during 8 months of culture. The alpha1 (II) procollagen gene is expressed only during the first 2 months of culture and after 3 months its expression is undetectable, which is consistent with its absence in adult articular cartilage. By Western blotting, Type II collagen protein had been synthesized and deposited in both the cell-associated and further-removed matrix compartments at 7 and 14 days of culture. These data indicate that chondrocytes cultured in alginate beads could be preserved for immunohistochemistry and ISH and that culture of human chondrocytes in alginate beads may serve as a good model for studying cartilage-specific phenotype as well as factors that influence cartilage matrix turnover.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0022-1554
pubmed:author
pubmed:issnType
Print
pubmed:volume
49
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1211-20
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:11561005-Adult, pubmed-meshheading:11561005-Aggrecans, pubmed-meshheading:11561005-Blotting, Western, pubmed-meshheading:11561005-Cartilage, Articular, pubmed-meshheading:11561005-Cells, Cultured, pubmed-meshheading:11561005-Chondrocytes, pubmed-meshheading:11561005-Extracellular Matrix Proteins, pubmed-meshheading:11561005-Humans, pubmed-meshheading:11561005-Immunohistochemistry, pubmed-meshheading:11561005-In Situ Hybridization, pubmed-meshheading:11561005-Infant, Newborn, pubmed-meshheading:11561005-Lectins, C-Type, pubmed-meshheading:11561005-Male, pubmed-meshheading:11561005-Matrix Metalloproteinase 3, pubmed-meshheading:11561005-Matrix Metalloproteinase 8, pubmed-meshheading:11561005-Procollagen, pubmed-meshheading:11561005-Proteoglycans, pubmed-meshheading:11561005-RNA, Messenger
pubmed:year
2001
pubmed:articleTitle
Gene expression by human articular chondrocytes cultured in alginate beads.
pubmed:affiliation
Department of Biochemistry, Rush Medical College at Rush-Presbyterian-St Luke's Medical Center, Chicago, Illinois 60612, USA. schubins@rush.edu
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't