Short-term microcultures of lymphocytes from Chinese hamster peripheral blood

Abstract

A microtechnique for the culture of Chinese Hamster lymphocytes is described using Cooke microtiter plates with 100,000 leucocytes in a culture volume of 0.1 ml and a culture time of three days. The culture media used were RPMI 1640 and Trowell T8 supplemented with 20% foetal calf serum (FCS) and 2 mu PHA. The cells were harvested with a Skatron cell culture harvester using glass fibre filters. Various technical aspects of the lymphocyte cultures from the Chinese Hamster are described. The relevance of changes in culture conditions to the variability of culture results was analysed for PHA and FCS concentrations, different culture media, cell concentration, vessel shape and culture duration.