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Title: Culture temperature affects human chondrocyte messenger RNA expression in monolayer and pellet culture systems
Authors: Ito, Akira  kyouindb  KAKEN_id  orcid (unconfirmed)
Nagai, Momoko
Tajino, Junichi
Yamaguchi, Shoki
Iijima, Hirotaka
Zhang, Xiangkai
Aoyama, Tomoki  kyouindb  KAKEN_id
Kuroki, Hiroshi  kyouindb  KAKEN_id
Author's alias: 青山, 朋樹
黒木, 裕士
Issue Date: 26-May-2015
Publisher: Public Library of Science
Journal title: PLoS ONE
Volume: 10
Issue: 5
Thesis number: e0128082
Abstract: Cell-based therapy has been explored for articular cartilage regeneration. Autologous chondrocyte implantation is a promising cell-based technique for repairing articular cartilage defects. However, there are several issues such as chondrocyte de-differentiation. While numerous studies have been designed to overcome some of these issues, only a few have focused on the thermal environment that can affect chondrocyte metabolism and phenotype. In this study, the effects of different culture temperatures on human chondrocyte metabolism- and phenotype-related gene expression were investigated in 2D and 3D environments. Human chondrocytes were cultured in a monolayer or in a pellet culture system at three different culture temperatures (32° C, 37° C, and 41° C) for 3 days. The results showed that the total RNA level, normalized to the threshold cycle value of internal reference genes, was higher at lower temperatures in both culture systems. Glyceraldehyde-3- phosphate dehydrogenase (GAPDH) and citrate synthase (CS), which are involved in glycolysis and the citric acid cycle, respectively, were expressed at similar levels at 32° C and 37° C in pellet cultures, but the levels were significantly lower at 41° C. Expression of the chondrogenic markers, collagen type IIA1 (COL2A1) and aggrecan (ACAN), was higher at 37° C than at 32° C and 41° C in both culture systems. However, this phenomenon did not coincide with SRY (sex-determining region Y)-box 9 (SOX9), which is a fundamental transcription factor for chondrogenesis, indicating that a SOX9-independent pathway might be involved in this phenomenon. In conclusion, the expression of chondrocyte metabolism-related genes at 32° C was maintained or enhanced compared to that at 37° C. However, chondrogenesis-related genes were further induced at 37° C in both culture systems. Therefore, manipulating the culture temperature may be an advantageous approach for regulating human chondrocyte metabolic activity and chondrogenesis.
Rights: © 2015 Ito et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
DOI(Published Version): 10.1371/journal.pone.0128082
PubMed ID: 26010859
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