Nanog maintains human chondrocyte phenotype and function in vitro

Hongjun Zheng; Francoise Gourronc; Joseph A Buckwalter; James A Martin

doi:10.1002/jor.20989

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Nanog maintains human chondrocyte phenotype and function in vitro

Journal article

Peer reviewed

Nanog maintains human chondrocyte phenotype and function in vitro

Hongjun Zheng, Francoise Gourronc, Joseph A Buckwalter and James A Martin

Journal of orthopaedic research, Vol.28(4), pp.516-521

04/2010

DOI: 10.1002/jor.20989

PMID: 19834952

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Abstract

Previous work showed that Nanog, a homeobox family transcription factor, maintains embryonic stem cell pluripotency, suggesting that it has a role in stabilizing cell phenotype. Human chondrocytes lose their phenotype and dedifferentiate after relatively few passages in culture, changes that may limit their value in restoring damaged articular cartilage. We hypothesized that Nanog could stabilize the phenotype of cultured human chondrocytes in long-term monolayer cultures. To test this hypothesis, the human Nanog gene was stably transduced into human chondrocytes using a retroviral vector. Chondrocyte-specific gene expression (collagen type II, aggrecan, cartilage link protein, and Sox9) was measured by reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR in monolayer cultured chondrocytes transduced with Nanog and in control chondrocytes transduced with empty vector. In vitro cartilage matrix protein formation by Nanog-transduced and control cells was compared using Safranin-O and immunofluorescence stains. We found that after 25 passages, Nanog-transduced chondrocytes maintained significantly higher expression of collagen type II, aggrecan, and cartilage link protein genes than controls. Under chondrogenic conditions, Nanog-transduced cells produced significantly more cartilage-specific matrix than control cells. These findings support the hypothesis that Nanog maintains the human chondrocyte phenotype and function after long-term monolayer culture. Preservation of the chondrocyte phenotype may improve the ability of cultured chondrocytes to repair or restore articular cartilage.

Chondrocytes - cytology

Homeodomain Proteins - metabolism

Humans

Glycoproteins - metabolism

Collagen Type II - metabolism

Matrilin Proteins

Chondrocytes - metabolism

Extracellular Matrix Proteins - metabolism

Biomarkers - metabolism

SOX9 Transcription Factor - metabolism

Gene Transfer Techniques

Nanog Homeobox Protein

Transduction, Genetic

Cartilage Oligomeric Matrix Protein

Extracellular Matrix Proteins - genetics

Cells, Cultured

Gene Expression Regulation

Proteoglycans - metabolism

Retroviridae

Aggrecans - metabolism

Reverse Transcriptase Polymerase Chain Reaction

Aggrecans - genetics

Collagen Type II - genetics

Homeodomain Proteins - genetics

Phenotype

Genetic Vectors

Cell Dedifferentiation - genetics

SOX9 Transcription Factor - genetics

Proteoglycans - genetics

Details

Title: Subtitle: Nanog maintains human chondrocyte phenotype and function in vitro
Creators: Hongjun Zheng - Department of Orthopaedics and Rehabilitation, 1182 ML, University of Iowa, 200 Hawkins Drive, Iowa City, Iowa 52242, USA
Francoise Gourronc
Joseph A Buckwalter
James A Martin
Resource Type: Journal article
Publication Details: Journal of orthopaedic research, Vol.28(4), pp.516-521
DOI: 10.1002/jor.20989
PMID: 19834952
NLM abbreviation: J Orthop Res
ISSN: 0736-0266
eISSN: 1554-527X
Publisher: United States
Grant note: name: Department of Orthopaedics and Rehabilitation at the University of Iowa
Language: English
Date published: 04/2010
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Microbiology and Immunology; Stead Family Department of Pediatrics; Pharmaceutical Sciences and Experimental Therapeutics; Orthopedics and Rehabilitation; Injury Prevention Research Center
Record Identifier: 9984040394402771

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