T cell medium is necessary to proliferate T cells. There are several T cell medium which is classified according to supplements and their origin source such as FBS, hPL, sera, and protein etc. Also, Medium is classified following types; serum-free(SF), xeno-free (XF), animal component-free (ACF), animal origin-free(AOF), and chemically defined(CD). In this study, we used a total of 4 types of T cell mediums such as two of SF/XF, one of ACF, and one of AOF. Culturing the T cells for 9 days and compared cell morphology, proliferation, and cell viability. On day 9, we analyzed the expression of subset markers(CD3, CD4, CD8), activation marker(CD25), differentiation marker(CD127, CDR7, CD45RA), and the secretion of cytokine(IFN-γ, perforin, and granzyme B). As a result, SF/XF medium is more degree of aggregation than ACF and AOF medium, and AOF medium is similar or higher expansion than SF/XF and ACF medium. CD4 marker shows a high expression in SF/XF medium, CD25 marker shows high expression in the order of AOF, ACF, and SF/XF medium. Naïve cells(CCR7+CD45RA+) are at the highest rate in ACF medium. IFN-γ secretion is lower in SF/XF mediums, and perforin secretion is lower in AOF medium. We conclude that AOF medium is similar to SF/XF medium in proliferation and similar to ACF medium in marker expression. This study suggests that T cell characteristics differ depending on T cell medium types, particularly, the origin source.