Low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes

J. Malda, Clemens van Blitterswijk, M. van Geffen, D.E. Martens, J. Tramper, J.U. Riesle

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Objective: To determine the effect of dissolved oxygen tension (DO) on the redifferentiation of dedifferentiated adult human nasal septum chondrocytes cultured as pellets. - Design: After isolation, human nasal chondrocytes were expanded in monolayer culture, which resulted in their dedifferentiation. Dedifferentiated cells were pelleted, transferred to a bioreactor and maintained for up to 21 days at 100% DO (21% oxygen), 25% DO (5.25% oxygen) or 5% DO (1% oxygen), which was controlled in the liquid phase. Redifferentiation was assessed by staining the extracellular matrix with safranin-O and by the immunolocalization of collagen types I, II, IX and of a fibroblast membrane marker (11-fibrau). In addition, glycosaminoglycans (GAG) and DNA content were determined spectrophotometrically. - Results: In monolayer culture, cells dedifferentiated and multiplied 90- to 100-fold. Cell pellets cultured in a bioreactor under conditions of low oxygen tension (25% DO or 5% DO) stained intensely for GAGs and for collagen type II, but very weakly for collagen type I. After 14 days of culturing, cell pellets maintained at 5% DO stained more intensely for collagen IX and more weakly for 11-fibrau than did those incubated at 25% DO. After 21 days of culturing the GAG content of cell pellets maintained at 5% DO was significantly greater than that of those incubated at 25% DO. Under air-saturated conditions (100% DO), the DNA and GAG contents of cell pellets decreased with time in culture. After 21 days of culturing, both parameters were substantially lower in cell pellets maintained at 100% DO than in those incubated at low oxygen tensions. The staining signals for collagen types II and IX were much weaker, and those for the markers of dedifferentiation (collagen type I and 11-fibrau) much stronger under air-saturated conditions than at low oxygen tensions. - Conclusion: These observations demonstrate that using the present set-up, low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes in pellet cultures.
Original languageUndefined
Pages (from-to)306-313
JournalOsteoarthritis and cartilage
Volume12
Issue number4
DOIs
Publication statusPublished - 2004

Keywords

  • Redifferentiation
  • Oxygen tension
  • IR-76257
  • dedifferentiation
  • METIS-237078
  • nasal septum chondrocytes

Cite this

Malda, J., van Blitterswijk, C., van Geffen, M., Martens, D. E., Tramper, J., & Riesle, J. U. (2004). Low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes. Osteoarthritis and cartilage, 12(4), 306-313. https://doi.org/10.1016/j.joca.2003.12.001
Malda, J. ; van Blitterswijk, Clemens ; van Geffen, M. ; Martens, D.E. ; Tramper, J. ; Riesle, J.U. / Low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes. In: Osteoarthritis and cartilage. 2004 ; Vol. 12, No. 4. pp. 306-313.
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abstract = "Objective: To determine the effect of dissolved oxygen tension (DO) on the redifferentiation of dedifferentiated adult human nasal septum chondrocytes cultured as pellets. - Design: After isolation, human nasal chondrocytes were expanded in monolayer culture, which resulted in their dedifferentiation. Dedifferentiated cells were pelleted, transferred to a bioreactor and maintained for up to 21 days at 100{\%} DO (21{\%} oxygen), 25{\%} DO (5.25{\%} oxygen) or 5{\%} DO (1{\%} oxygen), which was controlled in the liquid phase. Redifferentiation was assessed by staining the extracellular matrix with safranin-O and by the immunolocalization of collagen types I, II, IX and of a fibroblast membrane marker (11-fibrau). In addition, glycosaminoglycans (GAG) and DNA content were determined spectrophotometrically. - Results: In monolayer culture, cells dedifferentiated and multiplied 90- to 100-fold. Cell pellets cultured in a bioreactor under conditions of low oxygen tension (25{\%} DO or 5{\%} DO) stained intensely for GAGs and for collagen type II, but very weakly for collagen type I. After 14 days of culturing, cell pellets maintained at 5{\%} DO stained more intensely for collagen IX and more weakly for 11-fibrau than did those incubated at 25{\%} DO. After 21 days of culturing the GAG content of cell pellets maintained at 5{\%} DO was significantly greater than that of those incubated at 25{\%} DO. Under air-saturated conditions (100{\%} DO), the DNA and GAG contents of cell pellets decreased with time in culture. After 21 days of culturing, both parameters were substantially lower in cell pellets maintained at 100{\%} DO than in those incubated at low oxygen tensions. The staining signals for collagen types II and IX were much weaker, and those for the markers of dedifferentiation (collagen type I and 11-fibrau) much stronger under air-saturated conditions than at low oxygen tensions. - Conclusion: These observations demonstrate that using the present set-up, low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes in pellet cultures.",
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Malda, J, van Blitterswijk, C, van Geffen, M, Martens, DE, Tramper, J & Riesle, JU 2004, 'Low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes' Osteoarthritis and cartilage, vol. 12, no. 4, pp. 306-313. https://doi.org/10.1016/j.joca.2003.12.001

Low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes. / Malda, J.; van Blitterswijk, Clemens; van Geffen, M.; Martens, D.E.; Tramper, J.; Riesle, J.U.

In: Osteoarthritis and cartilage, Vol. 12, No. 4, 2004, p. 306-313.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes

AU - Malda, J.

AU - van Blitterswijk, Clemens

AU - van Geffen, M.

AU - Martens, D.E.

AU - Tramper, J.

AU - Riesle, J.U.

PY - 2004

Y1 - 2004

N2 - Objective: To determine the effect of dissolved oxygen tension (DO) on the redifferentiation of dedifferentiated adult human nasal septum chondrocytes cultured as pellets. - Design: After isolation, human nasal chondrocytes were expanded in monolayer culture, which resulted in their dedifferentiation. Dedifferentiated cells were pelleted, transferred to a bioreactor and maintained for up to 21 days at 100% DO (21% oxygen), 25% DO (5.25% oxygen) or 5% DO (1% oxygen), which was controlled in the liquid phase. Redifferentiation was assessed by staining the extracellular matrix with safranin-O and by the immunolocalization of collagen types I, II, IX and of a fibroblast membrane marker (11-fibrau). In addition, glycosaminoglycans (GAG) and DNA content were determined spectrophotometrically. - Results: In monolayer culture, cells dedifferentiated and multiplied 90- to 100-fold. Cell pellets cultured in a bioreactor under conditions of low oxygen tension (25% DO or 5% DO) stained intensely for GAGs and for collagen type II, but very weakly for collagen type I. After 14 days of culturing, cell pellets maintained at 5% DO stained more intensely for collagen IX and more weakly for 11-fibrau than did those incubated at 25% DO. After 21 days of culturing the GAG content of cell pellets maintained at 5% DO was significantly greater than that of those incubated at 25% DO. Under air-saturated conditions (100% DO), the DNA and GAG contents of cell pellets decreased with time in culture. After 21 days of culturing, both parameters were substantially lower in cell pellets maintained at 100% DO than in those incubated at low oxygen tensions. The staining signals for collagen types II and IX were much weaker, and those for the markers of dedifferentiation (collagen type I and 11-fibrau) much stronger under air-saturated conditions than at low oxygen tensions. - Conclusion: These observations demonstrate that using the present set-up, low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes in pellet cultures.

AB - Objective: To determine the effect of dissolved oxygen tension (DO) on the redifferentiation of dedifferentiated adult human nasal septum chondrocytes cultured as pellets. - Design: After isolation, human nasal chondrocytes were expanded in monolayer culture, which resulted in their dedifferentiation. Dedifferentiated cells were pelleted, transferred to a bioreactor and maintained for up to 21 days at 100% DO (21% oxygen), 25% DO (5.25% oxygen) or 5% DO (1% oxygen), which was controlled in the liquid phase. Redifferentiation was assessed by staining the extracellular matrix with safranin-O and by the immunolocalization of collagen types I, II, IX and of a fibroblast membrane marker (11-fibrau). In addition, glycosaminoglycans (GAG) and DNA content were determined spectrophotometrically. - Results: In monolayer culture, cells dedifferentiated and multiplied 90- to 100-fold. Cell pellets cultured in a bioreactor under conditions of low oxygen tension (25% DO or 5% DO) stained intensely for GAGs and for collagen type II, but very weakly for collagen type I. After 14 days of culturing, cell pellets maintained at 5% DO stained more intensely for collagen IX and more weakly for 11-fibrau than did those incubated at 25% DO. After 21 days of culturing the GAG content of cell pellets maintained at 5% DO was significantly greater than that of those incubated at 25% DO. Under air-saturated conditions (100% DO), the DNA and GAG contents of cell pellets decreased with time in culture. After 21 days of culturing, both parameters were substantially lower in cell pellets maintained at 100% DO than in those incubated at low oxygen tensions. The staining signals for collagen types II and IX were much weaker, and those for the markers of dedifferentiation (collagen type I and 11-fibrau) much stronger under air-saturated conditions than at low oxygen tensions. - Conclusion: These observations demonstrate that using the present set-up, low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes in pellet cultures.

KW - Redifferentiation

KW - Oxygen tension

KW - IR-76257

KW - dedifferentiation

KW - METIS-237078

KW - nasal septum chondrocytes

U2 - 10.1016/j.joca.2003.12.001

DO - 10.1016/j.joca.2003.12.001

M3 - Article

VL - 12

SP - 306

EP - 313

JO - Osteoarthritis and cartilage

JF - Osteoarthritis and cartilage

SN - 1063-4584

IS - 4

ER -