Effects of oxygen and culture system on in vitro propagation and redifferentiation of osteoarthritic human articular chondrocytes

Schrobback, Karsten, Klein, Travis Jacob, Crawford, Ross, Upton, Zee, Malda, Jos, & Leavesley, David Ian (2012) Effects of oxygen and culture system on in vitro propagation and redifferentiation of osteoarthritic human articular chondrocytes. Cell and Tissue Research, 347(3), pp. 649-663.

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Abstract

Regenerative medicine-based approaches for the repair of damaged cartilage rely on the ability to propagate cells while promoting their chondrogenic potential. Thus, conditions for cell expansion should be optimized through careful environmental control. Appropriate oxygen tension and cell expansion substrates and controllable bioreactor systems are probably critical for expansion and subsequent tissue formation during chondrogenic differentiation. We therefore evaluated the effects of oxygen and microcarrier culture on the expansion and subsequent differentiation of human osteoarthritic chondrocytes. Freshly isolated chondrocytes were expanded on tissue culture plastic or CultiSpher-G microcarriers under hypoxic or normoxic conditions (5% or 20% oxygen partial pressure, respectively) followed by cell phenotype analysis with flow cytometry. Cells were redifferentiated in micromass pellet cultures over 4 weeks, under either hypoxia or normoxia. Chondrocytes cultured on tissue culture plastic proliferated faster, expressed higher levels of cell surface markers CD44 and CD105 and demonstrated stronger staining for proteoglycans and collagen type II in pellet cultures compared with microcarrier-cultivated cells. Pellet wet weight, glycosaminoglycan content and expression of chondrogenic genes were significantly increased in cells differentiated under hypoxia. Hypoxia-inducible factor-3alpha mRNA was up-regulated in these cultures in response to low oxygen tension. These data confirm the beneficial influence of reduced oxygen on ex vivo chondrogenesis. However, hypoxia during cell expansion and microcarrier bioreactor culture does not enhance intrinsic chondrogenic potential. Further improvements in cell culture conditions are therefore required before chondrocytes from osteoarthritic and aged patients can become a useful cell source for cartilage regeneration.

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ID Code: 55590
Item Type: Journal Article
Refereed: Yes
Additional Information: Schrobback, Karsten
Klein, Travis Jacob
Crawford, Ross
Upton, Zee
Malda, Jos
Leavesley, David Ian
Germany
Cell Tissue Res. 2012 Mar;347(3):649-63. doi: 10.1007/s00441-011-1193-7. Epub 2011 Jun 4.
Keywords: Aged, Biological Markers/metabolism, Cartilage, Articular/*pathology, Cell Culture Techniques/*methods, Cell Differentiation/*drug effects/genetics, Cell Hypoxia/drug effects, Cell Proliferation/drug effects, Cells, Cultured, Chondrocytes/drug effects/*pathology, Chondrogenesis/drug effects/genetics, DNA/metabolism, Extracellular Matrix/drug effects/metabolism, Female, Gene Expression Regulation/drug effects, Glycosaminoglycans/metabolism, Humans, Hypoxia-Inducible Factor 1/metabolism, Immunohistochemistry, Immunophenotyping, Osteoarthritis/*pathology, Oxygen/*pharmacology, RNA, Messenger/genetics/metabolism
DOI: 10.1007/s00441-011-1193-7
ISSN: 1432-0878
Subjects: Australian and New Zealand Standard Research Classification > ENGINEERING (090000) > BIOMEDICAL ENGINEERING (090300)
Divisions: Current > Schools > School of Chemistry, Physics & Mechanical Engineering
Current > QUT Faculties and Divisions > Faculty of Health
Current > Institutes > Institute of Health and Biomedical Innovation
Current > QUT Faculties and Divisions > Science & Engineering Faculty
Copyright Owner: Copyright 2012 Springer
Copyright Statement: The original publication is available at SpringerLink
http://www.springerlink.com
Deposited On: 12 Dec 2012 22:57
Last Modified: 19 Dec 2012 10:17

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