Effects of Retinoic Acid and Valproic Acid on differentiation of Embryonic Neural Stem Cells

Abstract

Neural stem cells (NSC) have a myriad of potential uses, such as the treatment for spinal cord injury. Both retinoic acid (RA) and valproic acid (VPA) have been proven to be involved in neurogenesis in mice model; VPA as an HDAC inhibitor to induce neuronal differentiation and RA which enhances histone H3 acetylation to induce astrocyte differentiation of NSC.  The objective of this research is to find the most suitable substance and/or combination of substances to be used in neuronal differentiation of ESC-derived NSCs.

NS cells were derived first from mouse ES cells. DMEM Ham’s F-12 along with Vitamin B27, PSF, EGF, and BFGF was used. For experiment cells were moved to a four-well dish with N2 medium and FGF; control, RA, RA+FBS, RA+VPA. Immunocytostaining was done with antibodies for Tuj1, GFAP, and nestin, and then observed under fluorescent microscope. Cell count was done to determine differentiation of cells in each dish.

In treatment of the VPA and RA, there is indication that neurogenesis enhanced compared to cells that was only treated with RA and the control dish. The enhancement is mostly shown in the Tuj1 immunofluorescence where it is more abundant. However the number of astrocyte differentiation was also increased in the combination of RA and VPA treatment.

This indicates that there is no specific fate preference from the treatment. It does however indicate that during the incubation period there was an increase in cell proliferation and differentiation of NSCs when treated with a combination of VPA and RA.