TY - JOUR
T1 - Gene expression profiling of dedifferentiated human articular chondrocytes in monolayer culture
AU - Ma, Bin
AU - Leijten, Jeroen Christianus Hermanus
AU - Wu, Ling
AU - Kip, Michelle Maria Aleida
AU - van Blitterswijk, Clemens
AU - Post, Janine Nicole
AU - Karperien, Hermanus Bernardus Johannes
N1 - Open access
PY - 2013/1/30
Y1 - 2013/1/30
N2 - Objective
When primary chondrocytes are cultured in monolayer, they undergo dedifferentiation during which they lose their phenotype and their capacity to form cartilage. Dedifferentiation is an obstacle for cell therapy for cartilage degeneration. In this study, we aimed to systemically evaluate the changes in gene expression during dedifferentiation of human articular chondrocytes to identify underlying mechanisms.
Methods
RNA was isolated from monolayer-cultured primary human articular chondrocytes at serial passages. Gene expression was analyzed by microarray. Based on the microarray analysis, relevant genes and pathways were identified. Their functions in chondrocyte dedifferentiation were further investigated.
Results
In vitro expanded human chondrocytes showed progressive changes in gene expression. Strikingly, an overall decrease in total gene expression was detected, which was both gradual and cumulative. DNA methylation was in part responsible for the expression downregulation of a number of genes. Genes involved in many pathways such as the ERK and BMP pathways exhibited significant changes in expression. Inhibition of ERK pathway didn’t show dramatic effects in counteracting dedifferentiation process. BMP-2 was able to decelerate the dedifferentiation and reinforce the maintenance of chondrocyte phenotype in monolayer culture.
Conclusion
Our study not only improves our knowledge of the intricate signaling network regulating maintenance of chondrocyte phenotype, but also contributes to improved chondrocyte expansion and chondrogenic performance for cell therapy.
AB - Objective
When primary chondrocytes are cultured in monolayer, they undergo dedifferentiation during which they lose their phenotype and their capacity to form cartilage. Dedifferentiation is an obstacle for cell therapy for cartilage degeneration. In this study, we aimed to systemically evaluate the changes in gene expression during dedifferentiation of human articular chondrocytes to identify underlying mechanisms.
Methods
RNA was isolated from monolayer-cultured primary human articular chondrocytes at serial passages. Gene expression was analyzed by microarray. Based on the microarray analysis, relevant genes and pathways were identified. Their functions in chondrocyte dedifferentiation were further investigated.
Results
In vitro expanded human chondrocytes showed progressive changes in gene expression. Strikingly, an overall decrease in total gene expression was detected, which was both gradual and cumulative. DNA methylation was in part responsible for the expression downregulation of a number of genes. Genes involved in many pathways such as the ERK and BMP pathways exhibited significant changes in expression. Inhibition of ERK pathway didn’t show dramatic effects in counteracting dedifferentiation process. BMP-2 was able to decelerate the dedifferentiation and reinforce the maintenance of chondrocyte phenotype in monolayer culture.
Conclusion
Our study not only improves our knowledge of the intricate signaling network regulating maintenance of chondrocyte phenotype, but also contributes to improved chondrocyte expansion and chondrogenic performance for cell therapy.
KW - IR-84105
KW - METIS-294114
U2 - 10.1016/j.joca.2013.01.014
DO - 10.1016/j.joca.2013.01.014
M3 - Article
SN - 1063-4584
VL - 21
SP - 599
EP - 603
JO - Osteoarthritis and cartilage
JF - Osteoarthritis and cartilage
IS - 4
ER -