Adaptation of cell lines to suspension culture
AssigneeMonsanto Company
InventorsWilliam R. Tolbert, Joseph Feder
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Abstract
Densely packed cells growing rapidly in substrate-attached culture are gently released from the substrate by enzymatic treatment to form aggregates ranging from 20 .mu. to 500 .mu. in diameter. These aggregates are then transferred to fresh medium in agitated liquid suspension. The cells thereby continue to grow in aggregates whereby a normally anchorage-dependent estabished mammalian cell line is rapidly adapted to suspension culture.
Claims12
A method of rapidly adapting normally anchorage-dependent established cell lines to agitated liquid suspension culture comprising growing an inoculum of said cell line in substrate-attached culture to a stage beyond that of ordinary monolayer growth whereby multilayers of cells are produced, releasing the cells from the substrate-attached culture in the form of cell aggregates ranging from about 20 microns to about 500 microns in diameter, transferring said aggregates into a liquid suspension of nutrient culture medium of known essential amino acids, mineral salts, vitamins and carbohydrates, incubating at a temperature of from about 30.degree. C to about 38.degree. C while substantially continuously maintaining agitation of said medium, periodically transferring the growing cells into fresh nutrient medium and continuing the agitated incubation whereby said cells continue to be proliferated in aggregates.
Dependencies are inferred from the claim text.