Photosynthetic microorganisms produce relatively large amounts of physiologically active materials which stimulate the physiological activity of other organisms. In this study, mammalian HeLa cells were cultured in different culture media which were Dulbecco's modified Eagle medium (DMEM) with newborn calf serum (NCS), and DMEM including different types of physiologically activating compounds (PACs) extracted from Rhodobacter sphaeroides grown under various culture conditions. R. sphaeroides was grown under the following five different culture conditions: anaerobically in the light, anaerobically in the dark and treated with dimethyl sulfoxide, aerobically in the dark for 48 h, in the light for 48 h, and in the light for 24 h and changed after previous culturing in the dark for 24 h. The growth of HeLa cell was measured by cell counting using a hemocytometer, and the fluorescent intensities of cellular lysosomes were measured to check the level of cellular stress caused by adding PACs. The growth of HeLa cells cultured in DMEM with PACs extracted from R. sphaeroides aerobically grown under dark conditions was enhanced compared to that of cells grown with NCS. We also found that a high concentration of pigments such as bacteriochlorophylls and carotenoids and a high concentration of arginine produced by R. sphaeroides aerobically grown in the dark were implicated in increased growth of the HeLa cells. Therefore, our results suggest that PACs extracted from R. sphaeroides aerobically cultured in dark conditions can enhance the physiological activity of mammalian cells and serve as nontoxic and bioavailable materials.