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Dev Dyn. 2008 Oct;237(10):2705-15. doi: 10.1002/dvdy.21606.

Human primary corneal fibroblasts synthesize and deposit proteoglycans in long-term 3-D cultures.

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  • 1Departments of Biochemistry and Ophthalmology, Boston University School of Medicine, Boston, Massachusetts 02118, USA.

Abstract

Our goal was to develop a 3-D multi-cellular construct using primary human corneal fibroblasts cultured on a disorganized collagen substrate in a scaffold-free environment and to use it to determine the regulation of proteoglycans over an extended period of time (11 weeks). Electron micrographs revealed multi-layered constructs with cells present in between alternating parallel and perpendicular arrays of fibrils. Type I collagen increased 2-4-fold. Stromal proteoglycans including lumican, syndecan4, decorin, biglycan, mimecan, and perlecan were expressed. The presence of glycosaminoglycan chains was demonstrated for a subset of the core proteins (lumican, biglycan, and decorin) using lyase digestion. Cuprolinic blue-stained cultures showed that sulfated proteoglycans were present throughout the construct and most prominent in its mid-region. The size of the Cuprolinic-positive filaments resembled those previously reported in a human corneal stroma. Under the current culture conditions, the cells mimic a development or nonfibrotic repair phenotype.

Copyright (c) 2008 Wiley-Liss, Inc.

PMID:
18624285
[PubMed - indexed for MEDLINE]
PMCID:
PMC3760227
Free PMC Article

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