The effects of isolation on chondrocyte gene expression

Danika M. Hayman, Todd J. Blumberg, C. Corey Scott, Kyriacos A. Athanasiou

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Tissue engineering of articular cartilage usually requires the isolation and culture of chondrocytes. Previous studies have suggested that enzymatic isolation may alter the metabolic activity and growth rate of chondrocytes. This study examined the effects of 4 common isolation protocols on chondrocyte gene expression, morphology, and total cell yield immediately following the digest (t = 0) and after 2 culture periods (24 h and 1 week). Cartilage expiants were digested using 1 of 4 protocols: (1) 6-h collagenase digest, (2) 22-h collagenase digest, (3) 45-min trypsin digest followed by a 3-h collagenase digest, or (4) 1.5-h pronase digest followed by a 3-h collagenase digest. Gene expression levels for glyceraldehyde-3-phosphate dehydrogenase, type I collagen, type II collagen, aggrecan, superficial zone protein, matrix metalloproteinase-1, and tissue inhibitor of metalloproteinase-1 were measured at t = 0 h, 24 h, and 1 week using quantitative reverse transcriptase-polymerase chain reaction. In this study, cell yield was greatest for the 22-h collagenase and pronase-collagenase digests. However, the data indicate that a 6-h collagenase digest has the fewest gene expression changes compared to native cells. For tissue engineering, data from this study suggest that when cell yield is critical, a 22-h collagenase digest is preferable, but when obtaining cells closest to native chondrocytes is more desired, the 6-h collagenase digest is more beneficial.

Original languageEnglish (US)
Pages (from-to)2573-2581
Number of pages9
JournalTissue Engineering
Volume12
Issue number9
DOIs
StatePublished - Sep 2006
Externally publishedYes

Fingerprint

Collagenases
Chondrocytes
Gene expression
Matrix Metalloproteinase 13
Cartilage
Gene Expression
Tissue engineering
Collagen
Pronase
Tissue Engineering
Polymerase chain reaction
Matrix Metalloproteinase 8
Phosphates
Aggrecans
Matrix Metalloproteinase 1
Glyceraldehyde-3-Phosphate Dehydrogenases
Tissue Inhibitor of Metalloproteinase-1
Matrix Metalloproteinase Inhibitors
Collagen Type II
Tissue

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology
  • Biotechnology

Cite this

Hayman, D. M., Blumberg, T. J., Scott, C. C., & Athanasiou, K. A. (2006). The effects of isolation on chondrocyte gene expression. Tissue Engineering, 12(9), 2573-2581. https://doi.org/10.1089/ten.2006.12.2573

The effects of isolation on chondrocyte gene expression. / Hayman, Danika M.; Blumberg, Todd J.; Scott, C. Corey; Athanasiou, Kyriacos A.

In: Tissue Engineering, Vol. 12, No. 9, 09.2006, p. 2573-2581.

Research output: Contribution to journalArticle

Hayman, DM, Blumberg, TJ, Scott, CC & Athanasiou, KA 2006, 'The effects of isolation on chondrocyte gene expression', Tissue Engineering, vol. 12, no. 9, pp. 2573-2581. https://doi.org/10.1089/ten.2006.12.2573
Hayman DM, Blumberg TJ, Scott CC, Athanasiou KA. The effects of isolation on chondrocyte gene expression. Tissue Engineering. 2006 Sep;12(9):2573-2581. https://doi.org/10.1089/ten.2006.12.2573
Hayman, Danika M. ; Blumberg, Todd J. ; Scott, C. Corey ; Athanasiou, Kyriacos A. / The effects of isolation on chondrocyte gene expression. In: Tissue Engineering. 2006 ; Vol. 12, No. 9. pp. 2573-2581.
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