Abstract
Dopamine(DA)ergic differentiation from ES cells was induced by 5 steps in McKay,s method: stage 1, maintenance of ES cells; stage 2, formation of EBs; stage 3, selection of nestin-positive cells; stage 4, expansion of nestin (+) cells; stage 5, induction to DAergic neurons. To investigate whether physiological low oxygen found in development and cytokines expressed in the DA-depleted striatum increase the production of DA neurons from ES-derived neural progenitor cells (NPCs), NPCs were treated with cytokine mixtures (100 pg/ml IL-1β, 1 ng/ml IL-11, 1 ng/ml LIF, 1 ng/ml GDNF) or lowered O2 (3.5%) on stage 4 and stage 5, followed by tyrosine hydroxylase (TH) immunostaining. Low oxygen from stage 4 increased total number of TH(+) cells (1.78-fold) and number of TH(+) cells per sphere (1.58-fold of control ) as compared to normal O2. Cytokine mixture significantly increased TH(+) cells (2.13-fold) compared to non-treated control. IL-1β during stage 4 exhibited major contribution in the effect of cytokine mixtures. Data suggest that physiologically relevant low oxygen in development and cytokines and trophic factors that were enhanced in injured brain cause in increase of DAergic neurons from ES-derived NSCs. [J Physiol Sci. 2006;56 Suppl:S212]
