Cryopreservation in Hybridoma Production

Abstract

Hybridomas are normally made by random fusion of immune spleen cells with myeloma cells. Selection of hybridomas that produce antibodies with the specificity one is looking for only occurs after establishment and growth of all obtained hybridomas. Depending on the purity and nature of the antigen used for immunization, a variable number of different screening tests must be performed before the specificities of the obtained hybridomas are defined. Since hybridomas grow quickly it is generally accepted that at least the first screening tests should be performed rapidly. This poses a problem when time-consuming screening protocols are deemed necessary. Furthermore the propagation and eventual preservation of large numbers of hybridoma cultures, which are likely to include many useless ones, involves much labor, considerable costs, and infection hazards. To cope with the first problem and to minimize the latter, cryopreservation of hybridomas can be applied. The technique to cryopreserve tissue cultured cells was developed some decades ago (1,2). Depending on their tissue origin, cultured cells proved to be susceptible to cryopreservation to varying degrees (3). Cells of the hematopoietic lineage are especially vulnerable.