Journal
BIOTECHNOLOGY LETTERS
Volume 35, Issue 8, Pages 1233-1245Publisher
SPRINGER
DOI: 10.1007/s10529-013-1211-9
Keywords
Bioreactor; Human mesenchymal stem cells; Metabolite profiles microcarriers; Proliferation; Regenerative medicine bioprocessing; Spinner-flask
Categories
Funding
- Biotechnology and Biological Sciences Research Council (BBSRC
- UK)
- Bioprocessing Research Industries Club (BRIC)
- Engineering and Physical Sciences Research Council (EPSRC
- UK)
- Lonza GmbH (Cologne, Germany)
- Engineering and Physical Sciences Research Council [EP/H028277/1] Funding Source: researchfish
- EPSRC [EP/H028277/1] Funding Source: UKRI
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For the first time, fully functional human mesenchymal stem cells (hMSCs) have been cultured at the litre-scale on microcarriers in a stirred-tank 5 l bioreactor, (2.5 l working volume) and were harvested via a potentially scalable detachment protocol that allowed for the successful detachment of hMSCs from the cell-microcarrier suspension. Over 12 days, the dissolved O-2 concentration was > 45 % of saturation and the pH between 7.2 and 6.7 giving a maximum cell density in the 5 l bioreactor of 1.7 x 10(5) cells/ml; this represents > sixfold expansion of the hMSCs, equivalent to that achievable from 65 fully-confluent T-175 flasks. During this time, the average specific O-2 uptake of the cells in the 5 l bioreactor was 8.1 fmol/cell h and, in all cases, the 5 l bioreactors outperformed the equivalent 100 ml spinner-flasks run in parallel with respect to cell yields and growth rates. In addition, yield coefficients, specific growth rates and doubling times were calculated for all systems. Neither the upstream nor downstream bioprocessing unit operations had a discernible effect on cell quality with the harvested cells retaining their immunophenotypic markers, key morphological features and differentiation capacity.
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