Guided differentiation of mouse embryonic stem cell

The mouse embryonic stem cell’s differentiation was guided by several treatments, and each stage of differentiation was examined. Keywords: development stage EBs were dissociated and plated onto tissue culture dishes in the serum-free medium containing Insulin/Transferrin/Selenium/Fibronectin (ITSF). After 6 days, the cells were expanded and plated onto polyornithine (15 mg ml-1) and laminin-coated plates (Becton Dickinson Labware, Bedford, MA) with the N2 medium (Johe, 1996) supplemented with laminin, bFGF (R&D Systems, Minneapolis, MN), murine N-terminal fragment of SHH and murine FGF8 isoform b (R&D Systems) (stage 4) . Finally the cells were induced to differentiated into DA neurons by removal of bFGF. The differentiation medium consisted of N2 medium supplemented with laminin in the presence or absence of cAMP, AA (Sigma, St. Louis, MO). The cells were incubated under differentiation conditions for 6–15 days.