Cloning hybridomas in a reversible three-dimensional alginate matrix.
Alginate is a transparent polymer of guluronic and mannuronic acids that provide a favorable microenvironment for cell growth. Alginate gelation is calcium dependent and temperature independent. To facilitate the isolation of stable and productive antibody-producing hybridomas, we have developed a technique of cloning hybridomas in the three-dimensional alginate matrix. To provide cavities for hybridoma growth, we encorporated 10-15% (v/v) gelatin into the alginate prior to gelation. We have cloned more than 90 monoclonal antibody-producing hybridomas using the alginate matrix. The alginate matrix is readily reversible with the addition of a calcium chelator. The alginate matrix permits efficient cloning in limited incubator space, without the use of a feeder layer, and with minimal amount of medium. The transparent matrix also permits easy screening for clonality and growth.
Li
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Division of Thoracic Surgery, Brigham and Women's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115.
Abdi
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Mentzer
S J
SJ
D000464:Alginates; D000818:Animals; D003470:Culture Media; D003584:Cytological Techniques; D005069:Evaluation Studies as Topic; D005782:Gels; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006825:Hybridomas / Q000166:cytology*; D051379:Mice
D000464:Alginates; D003470:Culture Media; D005782:Gels; D006603:Hexuronic Acids; D020723:Glucuronic Acid