An investigation of a variety of different separation procedures and culture conditions was made in order to evaluate the methods for culturing leucocytes in vitro of Syrian and Chinese hamsters. Dextran (6%), human AB serum and fetal calf serum (FCS) were used as separating agents. In the Chinese hamster the best results were obtained with AB serum whereas in the Syrian hamster adequate separation was obtained with all three agents. Supplementation of the culture medium with FCS provided the most favourable conditions for cell survival in culture. Maximal numbers of mitoses were obtained after 4 days of culture at 37 °C with the Syrian hamster (0.5%) and after 5 days with Chinese hamster (0.2%). Leucocytes of Syrian and Chinese hamsters were also cultured following immunization with each of the ABO sera as well as with FCS. Both macro- and micromethods were partially successful. Using the macromethod, no mitoses were observed in the Chinese hamster, but a few in Syrian hamsters immunized with B serum. With the micromethod, mitotic figures were observed in Syrian hamsters immunized with B serum, O serum and FCS, and Chinese hamsters with O serum and FCS. Leucocytes of the Chinese hamster proved difficult to culture successfully by any of the methods used, whereas, with the Syrian hamster, immunization with FCS and culture using micromethod gave the best results.
Separating Agents in Azeotropic Distillation of Sec-Butanol and Water
