Aged black garlic (ABG) was extracted with 20% ethanol and water (crude extracts) and fractionated into three categories (>10, 3-10, and <3 kDa). The effect of crude extract supplements on anti-My-10 hybridoma cell growth and IgG1 antibody production was investigated in suspension culture with a chemically defined protein-free medium. We observed that supplementation of ABG to the cell culture medium stimulated anti-My-10 hybridoma cell growth and production of IgG1 antibody, particularly with fractionated ABG of low molecular weight. The stimulation depended upon the concentration and the size of the fractionated ABG. We also found that the growth-promoting activity was not correlated with high antibody production. These results suggest that fractionated ABG is a novel and promising alternative as an animal cell culture supplement.