Biotechnology Letters

, 33:2325

Expansion of human mesenchymal stem cells on microcarriers

  • Christopher J. Hewitt
  • Ken Lee
  • Alvin W. Nienow
  • Robert J. Thomas
  • Mark Smith
  • Colin R. Thomas
Original Research Paper

DOI: 10.1007/s10529-011-0695-4

Cite this article as:
Hewitt, C.J., Lee, K., Nienow, A.W. et al. Biotechnol Lett (2011) 33: 2325. doi:10.1007/s10529-011-0695-4

Abstract

The effects on human mesenchymal stem cell growth of choosing either of two spinner flask impeller geometries, two microcarrier concentrations and two cell concentrations (seeding densities) were investigated. Cytodex 3 microcarriers were not damaged when held at the minimum speed, NJS, for their suspension, using either impeller, nor was there any observable damage to the cells. The maximum cell density was achieved after 8–10 days of culture with up to a 20-fold expansion in terms of cells per microcarrier. An increase in microcarrier concentration or seeding density generally had a deleterious or neutral effect, as previously observed for human fibroblast cultures. The choice of impeller was significant, as was incorporation of a 1 day delay before agitation to allow initial attachment of cells. The best conditions for cell expansion on the microcarriers in the flasks were 3,000 microcarriers ml−1 (ca. 1 g dry weight l−1), a seeding density of 5 cells per microcarrier with a 1 day delay before agitation began at NJS (30 rpm), using a horizontally suspended flea impeller with an added vertical paddle. These findings were interpreted using Kolmogorov’s theory of isotropic turbulence.

Keywords

Human mesenchymal stem cellsMicrocarriersRegenerative medicine bioprocessingSpinner flasks

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Christopher J. Hewitt
    • 1
  • Ken Lee
    • 1
    • 2
  • Alvin W. Nienow
    • 1
    • 2
  • Robert J. Thomas
    • 1
  • Mark Smith
    • 3
    • 4
  • Colin R. Thomas
    • 2
  1. 1.Department of Chemical Engineering, Centre for Biological EngineeringLoughborough UniversityLeicestershireUK
  2. 2.School of Chemical EngineeringUniversity of BirminghamBirminghamUK
  3. 3.Smith and Nephew, York Science ParkYorkUK
  4. 4.AedStem Ltd, BioCentre York Science ParkYorkUK