Attempts at differentiation towards a dopaminergic phenotype.

<p><b>A:</b> Conditions resulting in high production of TH were 1% FBS in ‘ITS’ on a substrate of Collagen IV where almost 40% of cells were neurons (NF200+) expressing both TH and DT. However these were not really mature cells as the cultures still expressed a high level of the stem cell marker Oct 4 (although Nestin and Musashi were reduced to about 20%). <b>B:</b> Highest percentage of neurons (expressing NF) as well as low Oct4 as measures of cellular maturity were attained with 5% FBS, on PLO/Laminin substrate grown directly from neurospheres. FBS5, 5% FBS; FBS1: 1%FBS; X: no FBS; GDNF: 25 ng/mL GDNF. <b>C</b>: Highest percentage of TH expressing cells was obtained with 3.5% FBS, on PLO/Laminin substrate grown from adherent stem cells. Three cultures were assessed for each condition. SVZ scs: stem cells derived from SVZ; SVZ diff: cells subjected to putative differentiation conditions. (Error bars: +SEM; n = 3). <b>D:</b> Western blot confirms protein expression of TH in ‘differentiated’ cells derived from four region sources of stem cell cultures from one human. 40 µg total protein was loaded in each lane.