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Arthritis Research and Therapy

, 13:R64

First Online: 15 April 2011Received: 26 July 2010Revised: 15 February 2011Accepted: 15 April 2011

Abstract

IntroductionMesenchymal stem cells MSC are highly attractive for use in cartilage regeneration. To date, MSC are usually recruited from subchondral bone marrow using microfracture. Recent data suggest that isolated cells from adult human articular cartilage, which express the combination of the cell-surface markers CD105 and CD166, are multi-potent mesenchymal progenitor cells MPC with characteristics similar to MSC. MPC within the cartilage matrix, the target of tissue regeneration, may provide the basis for in situ regeneration of focal cartilage defects. However, there is only limited information concerning the presence-abundance of CD105-CD166 MPC in human articular cartilage. The present study therefore assessed the relative percentage and particularly the zonal distribution of cartilage MPC using the markers CD105-CD166.

MethodsSpecimens of human osteoarthritic OA; n = 11 and normal n = 3 cartilage were used for either cell isolation or immunohistochemistry. Due to low numbers, isolated cells were expanded for 2 weeks and then analyzed by flow cytometry FACS or immunofluorescence in chamber slides for the expression of CD105 and CD166. Following immunomagnetic separation of CD166- OA cells, multi-lineage differentiation assays were performed. Also, the zonal distribution of CD166 cells within the matrix of OA and normal cartilage was analyzed by immunohistochemistry.

ResultsFACS analysis showed that 16.7 ± 2.1% mean ± SEM of OA and 15.3 ± 2.3 of normal chondrocytes n.s. were CD105-CD166 and thus carried the established MPC marker combination. Similarly, 13.2% ± 0.9% and 11.7 ± 2.1 of CD105-CD166cells, respectively, were identified by immunofluorescence in adherent OA and normal chondrocytes. The CD166 enriched OA cells showed a stronger induction of the chondrogenic phenotype in differentiation assays than the CD166 depleted cell population, underlining the chondrogenic potential of the MPC. Strikingly, CD166 cells in OA and normal articular cartilage sections 22.1 ± 1.7% and 23.6% ± 1.4%, respectively; n.s. were almost exclusively located in the superficial and middle zone.

ConclusionsThe present results underline the suitability of CD166 as a biomarker to identify and, in particular, localize and-or enrich resident MPC with a high chondrogenic potential in human articular cartilage. The percentage of MPC in both OA and normal cartilage is substantially higher than previously reported, suggesting a yet unexplored reserve capacity for regeneration.

AbbreviationsDMEM-F12 NutmixDulbecco-s modified Eagle-s medium-F12 Nutmix

FACSfluorescence-activated cell sorting

FCSfetal calf serum

IgGimmunoglobulin G

MPCmesenchymal progenitor cell

MSCmesenchymal stem cell

OAosteoarthritis

PBSphosphate-buffered saline

TGFtransforming growth factor.

Electronic supplementary materialThe online version of this article doi:10.1186-ar3320 contains supplementary material, which is available to authorized users.

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Autor: David Pretzel - Stefanie Linss - Steffen Rochler - Michaela Endres - Christian Kaps - Saifeddin Alsalameh - Raimund W Kinn

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







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