Semi-solid methylcellulose-based hybridoma selection and cloning medium without HAT (serum-containing).

• Individual cells are suspended in viscous medium and form physically separated, discrete colonies that are easily isolated
• Monoclonal cell lines are isolated in less time using fewer resources compared with selection and cloning by limiting dilution
• Diverse clones with a wide range of growth rates and productivities form discrete colonies in the viscous medium
• Rare and high-producing clones can be individually isolated more easily using simultaneous selection and cloning in semi-solid medium compared with selection in bulk liquid cultures

ClonaCell™‐HY Medium D without HAT is a semi-solid methylcellulose-based medium that does not contain any selection agents such as HAT (hypoxanthine, aminopterin, and thymidine), enabling alternative hybridoma or myeloma selection methods to be used. This product can be used to select and clone myelomas or hybridomas in one step by adding an appropriate selection agent to the medium. Individual parental clones and their progeny remain localized together in the semi-solid matrix as they grow to form distinct colonies. This prevents the loss of rare clones by overgrowth from faster-growing cells, as can occur during selection in a liquid medium, and facilitates the isolation of monoclonal colonies.

The colonies can be easily picked from the semi-solid medium by manual or robotic methods and dispersed into a liquid medium for screening and expansion. This serum-containing medium has been verified for use in mouse and rat hybridoma development and reportedly is compatible for production and cloning of myelomas and/or hybridomas using lymphocytes from a variety of host animals including human, mouse, rat, and hamster.