Activation of bone marrow stem cell colonies in spleen by fermented soybeans AglyMax and ImmuSoy

Defatted soybean and soybean germ were fermented by Aspergillus sp. by a method based on traditional. Japanese fermentation technology. From this process we developed AglyMax, a product rich in isoflavone aglycones, and ImmuSoy, a fermentation culture made by a propriety kojic mold fermentation. Both products have an immunopotentiation activity in the intestinal tract. Their biological effect on bone marrow stem cells had not been elucidated and thus was examined in this study. Six-week-old male and female ICR/JCL mice purchased from Charles River Japan were used. For female mice, groups a and b were fed a normal diet and group c was fed a 1% ImmuSoy diet. For male mice, groups a and b were fed a normal diet; groups c and d were fed AglyMax at 30 mg/kg body weight and 100 mg/kg body weight, respectively; and group h was fed a 10% miso diet. After 3 wk all groups except group a were whole-body irradiated by 8 Gy of gamma-irradiation and isologous bone marrow nucleated cells (0.5 x 106 cells/0.5 mL) were injected intravenously into all experimental groups. One week later, the mice were killed, the macroscopic nodules on the surface of spleen were numerated under a microscope, and some nodules were studied histopathologically. Cells were stained with hematoxylin and eosin, iron staining was done with potassium ferrocyanide, and immuno-histochemical staining was done for monoclonal anti-CD34 and CD45 antibodies. For female mice, average spleen weight (relative to a 10-g body weight) was 53.0 mg in group a, 8.5 mg in group b, and 16.4 mg in group c, with significant differences between groups b and c. Similarly, average numbers of spleen colonies were 0 in group a, 1.6 in group b, and 18.6 in group c, with a significant increase in group c. For male mice, average spleen colony numbers were significantly increased in groups d and h compared with group b. Although prominent spleen colony foci were composed of lymphoblasts, none of the CD34 nor CD45 antibodies reacted to those colony foci. This evidence indicates that fermented soybean may contain factors that will potentiate bone marrow stem cell proliferation in spleen.