Thymocyte-conditioned medium clones

Thymocyte-conditioned medium (MTM), 44-45 for in vitro immunization, 9 use in cell fusions, 44 use in stabilizing hybridoma clones, 44... 

Micklem, L. R., McCann, M. C., and James, K. (1978) The use of rat mixed thymocyte-conditioned medium for hybridoma production, cloning and revival. J. Immunol. Methods 104, 81-86. 

Allogeneic mixed thymocyte medium (4) is a conditioned medium used to aid cells recover from the fusion process. It also encourages division of hybridoma cells after dilution cloning when colonies derived from single cells are required. Rat thymi are used for this purpose to minimise the number of animals required to produce adequate numbers of cells. The medium contains a number of helper cytokines produced by the thymocytes. It is important that the thymocytes used are derived from two different rat strains as this causes co-stimulation and enhanced cytokine secretion by the cells. Mixed thymocyte medium must always be diluted with RPMI 1640 medium containing 15% FBS and is usually used at a dilution of 10-15%. 

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