Life Technologies Corporation

Invitrogen Corporation

Richard M Fike

Paul J Price

Dale F Gruber

Shawn Barrett

David Epstein

Glenn Godwin

Stephen Gorfien

Joyce L Dzimian

Don McClure

David Judd

William Biddle

Zhou Jiang

Bruce Branchaud

James M Kubiak

William Whitford

Mwita Phelps

Paul Gulde

Mary Reynolds

Gabrielle Lorenzo

Barbara M Dadey

Richard Hassett

Thomas E Bulera

Stefan Weiss

Patrick Pomeroy

Neva Alles

Deirdre Eychner

Ryan Boniface

Joyce L Wolanske

Matthew Kahn

Andrew Campbell

David A Epstein

Stephen F Gorfien

Glenn P Godwin

Robert Radominski

Barbara Dadey

Bruce Brandchaud

Shawn Barett

Provided herein are, inter alia, methods for preparing a liquid cell culture media that has lesser lot-to-lot analytical variation, increased performance, and has lesser metal ion concentrations compared to a liquid media prepared by traditional methods. Such liquid media may be used for culturing cells, including but not limited to, recombinant cells.

Streamlined methods for making liquid media

Compositions and methods are described for preparing media, feeds, and supplements. Such methods and medias may display increased stability of labile components and may use, for example, microsuspension and/or encapsulation technologies, chelation, and optionally, coating and/or mixing the labile compounds with anti-oxidants. The compositions may withstand thermal and/or irradiation treatment and have reduced virus number. These techniques may result in product with extended shelf-life, extended release of their internal components into culture, or in product that can be added aseptically into a bioreactor using minimal volumes. The compositions and methods may optimize the bioproduction workflow and increase efficiency.

Cell culture media and methods

Methods for increasing the stability of, or protecting, labile components such as ethanolamine, growth factors, vitamins, etc., in compositions such as a cell culture medium. Stability of the labile compound is increased either, by derivatization of the labile compound with chemicals, or by sequestering the labile compound. Sequestering can be done either by encapsulation within a microcapsule, or by the use of sequestering agents. Encapsulation includes the encapsulation of dendrimers complexes of susceptible compounds within the microcapsule, thereby providing the controlled release of the susceptible compound that was protected. These methods may improve and extend storage conditions of compositions comprising the labile compounds, improve shipping and handling of compositions comprising the labile compounds, such as dry media formulations, at room temperature rather than at lower temperatures thereby decreasing shipping costs. Stabilization of labile compounds in compositions such as dry format media can be viewed as a contribution to green technology.

Compositions and methods for stabilizing susceptible compounds

The present invention is directed generally to dry cell culture media or feeds in pellet formats which can be reconstituted into liquid media for culturing cells in vitro. Each pellet composition may comprise the same or a different composition; for example, different vitamins, amino acids, buffers, trace salts, pH, iron chelators, etc. The invention also relates to methods of making dry cell culture media by altering ratios of different pellet compositions, or, methods of making modular dry cell culture media, or customizing media formulations for growing a cell type using pellets. According to the invention, media pellets may be easier to handle either before reconstitution, during shipping and handling; and/or during reconstitution. Media pellets may be used in any container like bags including sterile, single use bags for preparing media formulations. The invention also relates to kits and culture systems using media pellets.

Pellets used in cell culture and methods of making thereof

Methods for increasing the stability of, or protecting, labile components, such as ethanolamine, growth factors, vitamins, etc., in compositions such as a cell culture medium, are described. Stability of the labile compound is increased either, by derivatization or by sequestering the labile compound. Sequestering can be done either by encapsulation within a microcapsule or by the use of sequestering agents. Encapsulation can provide controlled release of the susceptible compound being protected. These methods may improve or extend the storage conditions, the shipping and handling of media compositions comprising the labile compounds at room temperature rather than at lower temperatures, thereby decreasing shipping costs.

The present invention provides a cell culture medium formulation that supports the in vitro cultivation, particularly in suspension, of mammalian cells, particularly epithelial cells and fibroblast cells, and methods for cultivating mammalian cells in suspension in vitro using these media. The media comprise a basal medium and a polyanionic or polyanionic compound, preferably a polysulfonated or polysulfated compound, and more preferably dextran sulfate. The present invention also provides chemically defined, protein-free eukaryotic cell culture media comprising an iron chelate and zinc, which is capable of supporting the growth (and particularly the high-density growth of mammalian cells) in suspension culture, increasing the level of expression of recombinant protein in cultured cells, and/or increasing virus production in cultured cells.

Serum-free mammalian cell culture medium, and uses thereof

The present invention provides a cell culture medium formulation that supports the in vitro cultivation, particularly in suspension, of mammalian cells, particularly epithelial cells and fibroblast cells, and methods for cultivating mammalian cells in suspension in vitro using these media. The present invention also provides chemically defined, protein-free eukaryotic cell culture media comprising an iron chelate and zinc, which is capable of supporting the growth (and particularly the high-density growth of mammalian cells) in suspension culture, increasing the level of expression of recombinant protein in cultured cells, and/or increasing virus production in cultured cells. The invention also provides kits for use in the cultivation of a mammalian epithelial cell that comprise one or more containers, wherein a first container contains the culture medium of the invention. These kits may further comprise one or more additional containers containing one or more supplements.

Methods and compositions involving nutritive media, media supplements, media subgroup and buffer formulations are provided. Powder buffer formulations that produce particular ionic and pH conditions upon reconstitution with a solvent are also disclosed. Methods of producing these media, media supplements, media subgroups and buffer formulations as well as kits and methods for cultivation of procaryotic and eukaryotic cells (including human cells) are also provided. Also disclosed are methods of producing sterile powdered media, media supplements, powdered growth factors, media subgroup and buffer formulations. Methods of sterilization include gamma irradiation. Methods for producing dry cell powders where spray-drying a cell suspension may be used are disclosed. Cells, media, media supplements, media subgroups and buffer powders produced by these methods are also disclosed.

Dry powder cells and cell culture reagents and methods of production thereof

The present invention relates generally to a method to reduce, substantially reduce, inactivate or eliminate adventitious agents and/or toxins in a sample, particuarly in nutritive media, media supplements, media subgroups and buffer formulations. Specifically, the present invention provides powdered nutritive media, media supplements and media subgroups produced by the methods of the invention, particuarly cell culture media supplements (including powdered sera such as powdered fetal bovine serum (FBS)). The invention further provides powdered buffer formulations produced by the methods of the invention. The invention also provides kits and methods for cultivation of prokaryotic and eukaryotic cells, particularly bacterial cells, yeast cells, plant cells and animal cells (including human cells) using these nutritive media, media supplements, media subgroups and buffer formulations. The invention also relates to methods for producing storage stable cells having reduced, substantially reduced or eliminated adventitious agents or toxins.

Methods for reducing adventitious agents and toxins and cell culture reagents produced thereby

The present invention relates generally to nutritive medium, medium supplement, media subgroup and buffer formulations. Specifically, the present invention provides powder nutritive medium, medium supplement and medium subgroup formulations, particularly cell culture medium supplements (including powdered sera such as powdered fetal bovine serum (FBS)), medium subgroup formulations and cell culture media comprising all of the necessary nutritive factors that facilitate the in vitro cultivation of cells. The invention further provides powder buffer formulations that produce particular ionic and pH conditions upon reconstitution with a solvent. The invention is particularly directed to methods of production of these media, media supplement, media subgroup and buffer formulations, and also provides kits and methods for cultivation of prokaryotic and eukaryotic cells, particularly bacterial cells, yeast cells, plant cells and animal cells (including human cells) using these dry powder nutritive media, media supplement, media subgroup and buffer formulations. The invention also relates to methods of producing sterile powdered media, media supplement (particularly powdered sera such as powdered FBS, powdered transferrin, powdered insulin, powdered organ extracts (such as bovine brain or pituitary extracts), powdered growth factors (such as EGF, FGF, etc.) and the like), media subgroup and buffer formulations. In a particularly preferred aspect, the invention relates to such methods wherein the sterilization is accomplished by gamma irradiation. The invention also relates to methods for producing dry cell powders, comprising spray-drying a cell suspension. The invention also relates to cell, media, media supplement, media subgroup and buffer powders produced by these methods.

Growing community of inventors

Clarence, NY, United States of America

Richard M Fike