Effect of Cell Concentrations and Culture Media on Hamster Splenocyte Viability During in vitro Cultivation
Keywords:
Hamster, Splenocyte, Immune cellsAbstract
Syrian golden hamster has been widely used as experimental model in several diseases, although their immune background still not fully understood due to the limited of antibodies in vitro cultivation of their immune cells. Therefore, knowledge of mouse immunity has often been substituted to explain the results of hamster experiments. However, as these immune systems have also been reported to be different., this methodology may be problematic. In order to study the hamster immunity, in vitro culture of hamster immune cells is required. In this study, we therefore proposed to optimize the conditions necessary to study the hamster splenic cells, which contains major source of immune cells. Normal hamster splenocytes at 1x105, 1x106, and 1x107 cells/mL were used to optimize culture conditions to obtain highest possible cell viability. Various media and supplements such as RPMI, DMEM, IMDM and 10-20% FBS were titrated. The cultures were performed over 1-4 days as has been done in most immunological studies. Our results demonstrated that after 2 days of culture, 1x107 cells/mL gave the maximum viabilities of 78.77% while 1x105 and 1x106 cells/mL concentration yielded 62.32% and 63.49% viability. After 4 days, the same 1x107 cells/mL also gave the maximum viabilities at 69.22%. A 15% FBS provided maximum viability of the 1x107 cells/mL concentration. RPMI medium provided an optimal viability of 70% after 4 days for the hamster splenic cells. RPMI and DMEM were subsequently deemed to be suitable for hamster splenocytes. This novel results for optimization of culture conditions will therefore be useful for study of hamster immune cells in other immunological techniques.
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