Test of a novel transparent floating carrier by recombinant cho cells expressing sIL6 receptor protein.
We describe a carrier with some unusual properties."Cellrafts" increase yields from adherent cells in conventional static T150 flasks, by floating multilayer strips of transparent film at the oxygen-rich surface of unstirred medium. This new technique allowed microscopic inspection of cell growth inside the carriers during bulk culture. Individual carriers could be picked out for subculture of selected colonies. A novel surface treatment by hypochlorite/uv allowed recycling of used carriers. Cellrafts' open-deck structure facilitated trypsinization with90% release as viable single cells from bulk carriers. Macro size (10mm by 1 mm) enables retention in flask by a coarse sieve insert in its neck, facilitating separation of product cells or media. Residual cells in carriers regenerated repeated harvests without need for reseeding. Carriers were tested with shear-sensitive CHO clones expressing soluble human IL6 receptor (sIL6R). Control was monolayer bulk culture on trays. Floating multilayer cultures remained viable longer than monolayers, had higher cellular activity of protein expression, and were less serum dependent (resembling cells on porous carriers). Purity and anti-sIL6R binding were identical to control product. Cellrafts were also tested in a small spinner vessel, but for litre batches this proved less convenient than in T-flasks. Though yields are low compared to well established porous carrier technology (spinner or packed bed) static transparent carriers might provide transitional scaleup from normal cytogenetics laboratory culture.
Gurari-Rotman
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Department of Molecular Genetics, Weizmann Institute of Science, 76 100, Rehovot, Israel.
Marks
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Chevath
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Revel
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Maroudas
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