Collagen type I and decorin expression in tenocytes depend on the cell isolation methodReportar como inadecuado




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BMC Musculoskeletal Disorders

, 13:140

First Online: 08 August 2012Received: 06 November 2011Accepted: 05 July 2012

Abstract

BackroundThe treatment of rotator cuff tears is still challenging. Tendon tissue engineering TTE might be an alternative in future. Tenocytes seem to be the most suitable cell type as they are easy to obtain and no differentiation in vitro is necessary. The aim of this study was to examine, if the long head of the biceps tendon LHB can deliver viable tenocytes for TTE. In this context, different isolation methods, such as enzymatic digestion ED and cell migration CM, are investigated on differences in gene expression and cell morphology.

MethodsSamples of the LHB were obtained from patients, who underwent surgery for primary shoulder arthroplasty. Using ED as isolation method, 0.2% collagenase I solution was used. Using CM as isolation method, small pieces of minced tendon were put into petri-dishes. After cell cultivation, RT-PCR was performed for collagen type I, collagen type III, decorin, tenascin-C, fibronectin, Scleraxis, tenomodulin, osteopontin and agreccan.

ResultsThe total number of isolated cells, in relation to 1 g of native tissue, was 14 times higher using ED. The time interval for cell isolation was about 17 hours using ED and approximately 50 days using CM. Cell morphology in vitro was similar for both isolation techniques. Higher expression of collagen type I could be observed in tenocyte-like cell cultures TLCC using ED as isolation method p < 0.05, however decorin expression was higher in TLCC using CM as isolation method p < 0.05. Dedifferentiation potential seemed to be similar for both isolation techniques.

ConclusionIn summary tenocyte-like cells can be obtained with both isolation methods ED and CM from the LHB. As no obvious disadvantage could be seen using ED, this method is more suitable for clinical use, as time for cell isolation is shorter and a remarkably higher number of cells can be obtained. However, both isolation methods can further be improved.

KeywordsTenocytes Tissue engineering Isolation method Gene expression AbbreviationsCMCell migration

EDEzymatic digestion

LHBLong head of the biceps tendon

TSPCTendon stem and progenitor cells

TLCCTenocyte-like cell culture

TTETendon tissue engineering.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2474-13-140 contains supplementary material, which is available to authorized users.

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Autor: Markus U Wagenhäuser - Matthias F Pietschmann - Birte Sievers - Denitsa Docheva - Matthias Schieker - Volkmar Jansson - P

Fuente: https://link.springer.com/







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