Gene expression during redifferentiation of human articular chondrocytes

Tallheden, Tommi; Karlsson, Camilla; Brunner, Andreas; Lee, Josefine van der; Hagg, Rupert; Tommasini, Roberto; Lindahl, Anders

doi:10.1016/j.joca.2004.03.004

Cited by 129 publications

(116 citation statements)

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“…These results are thus in accordance with previous results from our group. 11 In contrast to the chondrocytes, MSCs B differentiated towards a phenotype consisting of different types of cartilage. This was demonstrated by increased gene expression of markers for a hyaline (collagen type IIB), hypertrophic (collagen type X), and an immature (collagen type I) phenotype.…”

Section: Discussionmentioning

confidence: 99%

“…Chondrocytes were isolated by collagenase digestion overnight and expanded in DMEM/F12 (Invitrogen, Paisley, UK) supplemented with L-ascorbic acid (0.025 mg/mL; Apotekets Production Unit, Umeå , Sweden), gentamycin sulfate (50 mg/L; Gibco, Paisley, Scotland), amphoterricin B (250 mg/mL; Gibco), and L-glutamine (2 mM; Gibco) with addition of 10% human serum. 11 The above medium is referred to as Chondrocyte Culture Medium. MSCs isolated from the iliac crest of three donors (mean age 46 years, range 41-49) were purchased from Progentix (Bilthoven, Netherlands).…”

Section: Isolation and Culture Of Cellsmentioning

confidence: 99%

“…11 Briefly, 2 Â 10 5 cells were placed into a conical polypropylene tube with 0.5 mL of defined medium, consisting of DMEM-HG (PAA Laboratories, Linz, Austria) supplemented with 5.0 mg/mL linoleic acid (Sigma-Aldrich, Stockholm, Sweden), insulintransferrin-selenium-G (ITS-G) (Life Technologies, Paysley, UK), 1.0 mg/mL human serum albumin (Equitech-Bio, Kerrville, TX), 10 ng/mL transforming growth factor beta 1 (TGF-b1) (R&D Systems, Abingdon, UK), 10 À7 M dexamethasone (Sigma-Aldrich), 14 mg/mL ascorbate acid (Sigma-Aldrich), and 1% penicillinstreptomycin (PEST) (PAA Laboratories). The cells were centrifuged at 500Âg for 5 min and maintained at 378C in 7% CO 2 /93% air.…”

Section: Differentiation Modelmentioning

confidence: 99%

“…The sections were also subjected to immunohistochemistry as described previously. 11 Briefly, collagen type I and II were detected using monoclonal antibodies (ICN Biomedicals, Aurora, OH) and a secondary HRP-conjugated antibody (Jackson Immunoresearch Laboratories, West Grove, Pennsylvania). Endogenous peroxidase activity was quenched using 3% hydrogen peroxide.…”

Section: Differentiation Modelmentioning

confidence: 99%

“…11 Two pellets were separately analyzed per donor and time point for each of the three biochemical assays.…”

Section: Biochemical Analyses Of Pelletsmentioning

confidence: 99%

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Differentiation of human mesenchymal stem cells and articular chondrocytes: Analysis of chondrogenic potential and expression pattern of differentiation‐related transcription factors

Karlsson

Brantsing

Svensson

et al. 2006

Journal Orthopaedic Research

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Mesenchymal stem cells (MSCs) are a candidate for replacing chondrocytes in cellbased repair of cartilage lesions. However, it has not been clarified if these cells can acquire the hyaline phenotype, and whether chondrocytes and MSCs show the same expression patterns of critical control genes in development. In order to study this, articular chondrocytes and iliac crest derived MSCs were allowed to differentiate in pellet mass cultures. Gene expression of markers for the cartilage phenotype, helix-loop-helix (HLH) transcription factors, and chondrogenic transcription factors were analyzed by real-time PCR. Matrix production was assayed using biochemical analysis for hydroxyproline, glycosaminoglycans, and immunohistochemistry for collagen types I and II. Significantly decreased expression of collagen type I was accompanied by increased expression of collagen types IIA and IIB during differentiation of chondrocytes, indicating differentiation towards a hyaline phenotype. Chondrogenesis in MSCs on the other hand resulted in up-regulation of collagen types I, IIA, IIB, and X, demonstrating differentiation towards cartilage of a mixed phenotype. Expression of HES1 increased significantly during chondrogenesis in chondrocytes while expression in MSCs was maintained at a low level. The HLH gene HES5 on the other hand was only detected in chondrocytes. Expression of ID1 decreased significantly in chondrocytes while the opposite was seen in MSCs. These findings suggest that chondrocytes and MSCs differentiated and formed different subtypes of cartilage, the hyaline and a mixed cartilage phenotype, respectively. Differentially regulated HLH genes indicated the possibility for HLH proteins in regulating chondrogenic differentiation. This information is important to understand the potential use of MSCs in cartilage repair. ß

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Section: Discussionmentioning

confidence: 99%

Section: Isolation and Culture Of Cellsmentioning

confidence: 99%

Section: Differentiation Modelmentioning

confidence: 99%

Section: Differentiation Modelmentioning

confidence: 99%

“…11 Two pellets were separately analyzed per donor and time point for each of the three biochemical assays.…”

Section: Biochemical Analyses Of Pelletsmentioning

confidence: 99%

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Differentiation of human mesenchymal stem cells and articular chondrocytes: Analysis of chondrogenic potential and expression pattern of differentiation‐related transcription factors

Karlsson

Brantsing

Svensson

et al. 2006

Journal Orthopaedic Research

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Synthesis of monolithic HPLC stationary phase with self‐assembled molecular recognition sites for 4‐aminophenol

Baykara

Tunc

Ulubayram

2011

J of Applied Polymer Sci

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A molecularly imprinted polymer (MIP) monolith for selective recognition of 4-aminophenol (4-AMP) was prepared by in situ polymerization technique as high-performance liquid chromatography (HPLC) stationary phase. For this purpose, several 4-AMP imprinted monoliths were synthesized by using only methacrylic acid (MAA), acrylamide (AAM), or isobornyl methacrylate (IBMA) in the presence of high amount of crosslinker, ethylene glycol dimethacrylate (EDMA), and these polymeric monolith columns were connected to HPLC to evaluate their separation capabilities. By selection of appropriate functional monomer and optimization of polymerization conditions, MAA-based monolithic MIP showed good flow through properties, high selectivity to the templated molecule, and high resolution in the separation of paracetamol and its main impurity, 4-AMP. Besides, effective binding site density and dissociation constant of this monolith were estimated by using frontal chromatography and found as 7.95 lmol/g and 1.06 mM, respectively. Surface area of the same monolith was found as 23.48 m 2 /g from multipoint BET analysis.

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Epithelial Membrane Protein 1 Inhibits Human Spinal Chondrocyte Differentiation

Xiong

et al. 2011

The Anatomical Record

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The molecular mechanisms underlying human spinal chondrocyte differentiation remain unclear. We recently demonstrated that epithelial membrane protein 1 (EMP1) is highly expressed in degenerative intervertebral discs. EMP1 is involved in the differentiation of multiple cell types, including progenitor/pre-B cells, neurons, and podocytes. Therefore, we hypothesize that EMP1 may participate in the differentiation of spinal chondrocytes. We cultured chondrocytes from human nucleus pulposus. Through lentivirus-mediated knockdown and overexpression of EMP1, we find that EMP1 promotes cell proliferation and survival, alters cell morphology and cell cycle, reduces cell condensation, and inhibits cell hypertrophy and the expression of chondrocyte maturation markers such as collagen X, aggrecan, sex-determining region Y (SRY)-box 9, and runtrelated transcription factor 2. We also show that EMP1 is not expressed in the ossification center of vertebrae but is highly expressed in the nucleus pulposus and growth plate, where chondrocytes are immature and endochondral ossification has not occurred. These results suggest that EMP1 inhibits human spinal chondrocyte differentiation. Anat Rec, 294:1015Rec, 294: -1024Rec, 294: , 2011. V V C 2011 Wiley-Liss, Inc.

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Gene expression during redifferentiation of human articular chondrocytes

Abstract: Our data show that human articular chondrocytes taken from surplus cells of patient undergoing ACT treatment and expanded in ML, redifferentiate and form cartilage like matrix in vitro and that this dynamic process involves genes known to be expressed in early chondrogenesis.

Cited by 129 publications

References 58 publications

Differentiation of human mesenchymal stem cells and articular chondrocytes: Analysis of chondrogenic potential and expression pattern of differentiation‐related transcription factors

Differentiation of human mesenchymal stem cells and articular chondrocytes: Analysis of chondrogenic potential and expression pattern of differentiation‐related transcription factors

Synthesis of monolithic HPLC stationary phase with self‐assembled molecular recognition sites for 4‐aminophenol

Epithelial Membrane Protein 1 Inhibits Human Spinal Chondrocyte Differentiation

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