Abstract
Hybridoma and myeloma cell lines can be stored by slowly freezing cells in an appropriate solution of nutrients and a cryoprotectant such as glycerol or dimethyl sulfoxide (DMSO). In this protocol, cells are centrifuged at 4°C, resuspended in cold freezing solution (10% DMSO in FBS), and then transferred to an appropriate freezing vial. The vials are slowly frozen to −70°C in Styrofoam racks and then stored in liquid nitrogen (LN2). Cells stored in LN2 will remain viable for years. Once a frozen vial has been removed from LN2 storage, it should be thawed as described, grown out into log phase, and refrozen.
Publisher
Cold Spring Harbor Laboratory
Subject
General Biochemistry, Genetics and Molecular Biology
Cited by
1 articles.
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1. Growing Hybridomas;Cold Spring Harbor Protocols;2021-10