Bayer Pharmaceuticals Corporation

Mokhtar Mered

Rudiger Heidemann

Konstantin Konstantinov

William H Kelsey

De Qian Wang

James D Michaels

Bruce Gardner

A device and method for seed-train expansion of a source of mammalian cells and their use in production of proteins expressed by the cells comprising the use of a dedicated inoculation bioreactor for expanding the cells prior to the transfer to a production bioreactor is disclosed. The novel inoculation bioreactor is designed to facilitate an improved method of mammalian cell seed-train expansion, and is distinguished by the presence of an 'inoculation well' which communicates with the interior of the bioreactor and which facilitates the growth of mammalian cells for commercial seed-train expansion. The method comprises adding the cryopreserved cells to media within the inoculation well of the inoculation bioreactor, enabling the cells to grow to a predetermined concentration within the inoculation well by monitoring and adjusting the condition of the media and environment, and thereafter incrementally increasing the volume of the media within the reactor so that optimal cell growth is maintained.

Device and method for seed-train expansion of mammalian cells

The invention is a novel cell freezing and storage bag assembly and method for using the assembly in the seed train expansion of mammalian cells. The bag is constructed principally of polytetrafluoroethylene fabric. The bag is designed to hold enough cells that a bioreactor can be inoculated directly. The bag is designed to be filled to a fraction of its maximum capacity so that the cell suspension has a very thin cross-section (less than about 10 millimeters). The bag design includes a transfer set that can be sterilely welded to the source of the mammalian cells. This sterile-weldable transfer set allows the bags to be filled quickly with minimal risk of contamination. In the method, once each bag is filled, it is sealed below the connection with the transfer set and the bag is cut &ldquo;above&rdquo; the new seal (on the same side of the seal as the transfer set). When a bioreactor is to be inoculated, the contents of the bag are drained via a sterile-weldable inoculation line. During freezing and storage, the inoculation line is protected from mechanical damage.

Cryopreservation bag assembly for mammalian cell lines

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Orinda, CA, United States of America

Mokhtar Mered