Effects of retinoic acid and astrocyte as a feeder layer in differentiation of hippocampal stem cell

ABSTRACT

Adult stem cells are pluripotent cells conventionally isolated from some part of body by different methods. From developmental stand point, murine neural stem cells represent an accessible and important system for studies of basic stem cell property such as self- renewal and multipotency. In this study hippocampal stem cells obtained from embryonic day 18 [E18]. Pregnant female rats were killed, embryos heads separated and then hippocampus isolated by the method of Banker, then their cells dissociated by the methods of Fishbach, and plated in flask 25cm, after 3 days cells separated by tripsin, counted with trepan blue and hemocytometer, divided into two density [high 200000] and [low 20000 cells]. Before transplantation of cells, six well plates coated with poly L lysin and inactivated astrocyte. Then isolated cells transplanted into 6 well plate for 4 days with medium DMEM/F12 supplemented with FBS10%. After 4 days different doses of ARTA and RA cis-9 added per well for 6 days, and then immunocytochemistery were done. After 6 days of treatment with above factors, doses of 100nM RA cis-9 and 500 nM ATRA have the more staining cells with monoclonal antibody. But in 100 nM RA cis-9, we saw maximum differentiated cells. All of differentiation were done on wells with inactivated astrocyte layer in high and low density. Inactivated astrocyte as a feeder layer and extrinsic factors such as All Transe Retinoic Acid [ATRA] and RA cis-9 can cause differentiation in hippocampal stem cells into photoreceptor like cell