Optimization of physical parameters for cell attachment and growth on macroporous microcarriers

Autor: Michael Butler, J. M. Berry, Y C Ng
Rok vydání: 1996
Předmět:
Zdroj: Biotechnology and Bioengineering. 50:627-635
ISSN: 1097-0290
0006-3592
DOI: 10.1002/(sici)1097-0290(19960620)50:6<627::aid-bit3>3.0.co;2-m
Popis: The rates of cell attachment of the anchorage-dependent mammalian cell line Vero to the gelatin-based macroporous microcarrier Cultispher-G were determined under various conditions. An optimal rate of attachment (0.98 x 10(-2) min(-1)) occurred by an intermittent stirring regimen of 3 min stirring at 40 rpm per 33 min. This stirring regimen appeared to maximize cell-to-bead attachment and minimized cell aggregation which occurred at a broadly comparable rate.A further increase in the rate of cell-to-bead attachment occurred by preincubation of the microcarriers in serum-supplemented medium prior to cell inoculation in a serum-free medium. However, serum supplementation (>5%) was required for maximal cell growth. The pH of the medium had little effect on cell attachment over a broad range (pH 7.1-8.0). An initial cell/bead inoculum of 30 ensured an even distribution of cells on the available microcarriers with a low proportion of unoccupied beads.The rate of cell attachment to Cultispher-G was an order of magnitude lower than the determined value for the charged dextran microcarrier Cytodex-1, which was measured as 9.05 x 10(-2) min(-1). The optimal conditions for cell attachment were significantly different for the two bead types. Cell attachment to the electrostatic surface of the Cytodex-1 microcarriers was highly dependent on pH and serum supplementation. Cell aggregation during attachment to the Cytodex-1 microcarriers was minimal because of the higher rate of cell-microcarrier attachment.The porous nature of the Cultispher-G microcarriers allowed a maximum cell/bead loading of >1400, which was at least 3 times higher than equivalent loading of the cells on Cytodex-1. The Cultispher-G matrix also allowed the use of higher agitation rates (up to 100 rpm) in spinner flasks without affecting the cell growth rate or maximum cell density.
Databáze: OpenAIRE
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