The influence of nutrient supply and cell density on the growth and survival of intervertebral disc cells in 3D culture


The adult human intervertebral disc (IVD) is normally avascular. Changes to the extracellular matrix in degenerative disc disease may promote vascularisation and subsequently alter cell nutrition and disc homeostasis. This study examines the influence of cell density and the presence of glucose and serum on the proliferation and survival of IVD cells in 3D culture. Bovine nucleus pulposus (NP) cells were seeded at a range of cell densities (1.25 × 10(5)-10(6) cells/mL) and cultured in alginate beads under standard culture conditions (with 3.15 g/L glucose and 10 % serum), or without glucose and/or 20% serum. Cell proliferation, apoptosis and cell senescence were examined after 8 days in culture. Under standard culture conditions, NP cell proliferation and cluster formation was inversely related to cell seeding density, whilst the number of apoptotic cells and enucleated "ghost" cells was positively correlated to cell seeding density. Increasing serum levels from 10% to 20% was associated with increased cluster size and also an increased prevalence of apoptotic cells within clusters. Omitting glucose produced even larger clusters and also more apoptotic and senescent cells. These studies demonstrate that NP cell growth and survival are influenced both by cell density and the availability of serum or nutrients, such as glucose. The observation of clustered, senescent, apoptotic or "ghost" cells in vitro suggests that environmental factors may influence the formation of these phenotypes that have been previously reported in vivo. Hence this study has implications for both our understanding of degenerative disc disease and also cell-based therapy using cells cultured in vitro.

Divisions: College of Health & Life Sciences
College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine
College of Health & Life Sciences > School of Biosciences
Additional Information: © The author
Uncontrolled Keywords: animals,apoptosis,caattle,cell aging,cell count,cell culture techniques,cell proliferation,cell survival,food,glucose,intervertebral disc,serum,Biochemistry,Cell Biology,Bioengineering,Biomedical Engineering,Biomaterials,Medicine(all)
Publication ISSN: 1473-2262
Last Modified: 03 Jan 2024 18:36
Date Deposited: 29 May 2012 12:29
Full Text Link: http://www.ecmj ... pdf/v022a08.pdf
Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2011-12
Authors: Stephan, Simon
Johnson, W. Eustace
Roberts, Sally



Version: Published Version

License: Creative Commons Attribution

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