Promotion of proliferation of murine hematopoietic stem cells by growth factors in murine amniotic fluid

It has been shown that murine amniotic fluid (MAF) displays immunosuppressive activities. We examined the effect of MAF obtained from normal mice on a murine fetal liver cell (FLC) primary culture in vitro for the detection of the possible existence of cytokines that affect hematopoiesis. MAF promoted proliferation of murine FLC and;adult bone marrow cells. Proliferation-promoting activity of MAF was observed throughout the period between days 12 and 15 of pregnancy, peaking at day 12. Pooled MAF was subjected to purification procedures to isolate the active molecule(s). After ion exchange and size exclusion chromatography, one of the active molecules was shown to react with anti-mouse stem cell growth factor (SCF) antibody and the molecular weight was determined as 40-45 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. The proliferation-promoting activity of MAF was partially neutralised by the anti-SCF antibody. These results provide evidence that MAF may contain multiple growth factors for FLC, one of which may be a unique molecule related to SCF.