Third trimester NG2+ amniotic fluid cells respond to inflammation and are effective in SCI by means of HGF production amniotic fluid ameliorate the functional recovery in a mouse model of spinal cord injury

Aims Cellular treatment of spinal cord injury (SCI) has become a promising strategy. In a recent work (Bottai et al, 2014, Experimental Neurology in press) we reported that administration of at term Amniotic Fluid Cells (AFCs) improved recovery of motor function after SCI possibly through the enhancement of Hepatocyte Growth Factor (HGF) production. This report is about our attempt to depict the mechanism by which these cells exert their therapeutical effects in SCI that appear relate to the expression of the surface marker chondroitin sulfate proteoglycan (NG2). Methods The experimental plan is as follow: Cell cultures that have effectiveness and non effectiveness in our mouse model of SCI are plated in a multiwell plate. The following day, the medium is changed with a medium containing IL1β or Lipopolysaccharide or TNF-α. Cells are maintained in these media for various time points, then fixed. The fixed cells are stained for HGF antibody in order to evaluate the protein level expression in both the type of cells and the specific untreated controls. The expression of NG2 is knockdown by means of siRNA, and then the experiment as described in point 1 is carried out. Results We have previously shown that AFCs can increase their production of HGF when exposed to inflammatory agents. Preliminary data indicate that NG2 levels of expression could modulate HGF production. Conclusions This study can demonstrate that NG2 expression may correlate with HGF production in AFCs in vivo and in vitro.