Wichterle H, Lieberam I, Porter JA, Jessell TM. Directed differentiation of embryonic stem cells into motor neurons. Cell. 2002 Aug 9;110(3):385-97. PubMed Abstract

REAGENTS/MATERIAL:

Embryoinic stem cells cultured and embryoid bodies (EB) treated to retonic acid to develop neurons.

Immunocytochemistry EBs processed for immunohistochemistry with antibodies noted below.Images were obtained using a BioRad confocal microscope. Antibody Reagents:Rabbit aHB9, aOlig2, aNkx2.2, aLhx3, aDbx1 (Pierani, 1999), aIrx3, aGFP (1:2000, Molecular Probes), aSox1 (1:500 [Pevny et al., 1998]), aOtx2 (1:1000 [Mallamaci et al., 1996]), aHoxC5 (Liu et al., 2001), and aChx10; mouse aPax7, aPax6, aHB9, aLim1, aNkx6.1, aNkx2.2, aEn1, aLim2 (mouse specific), aNCAM (mouse specific), aSC1 (chick specific), aNeuN (1:500, Chemicon), aTuj1 (1:2000, BABCO), aSyb (mouse specific, 1:1000, Synaptic Systems), aSyn (1:1000, Synaptic Systems), and aNeurofilament; guinea pig a-Isl1, anti-Olig2, and anti-Hoxc6 (Liu et al., 2001); goat aChAT (1:200, Chemicon) and aVAChT (mouse specific, 1:1000, Chemicon) All antibodies cited in Novitch et al. (2001) when not otherwise specified. FITC-, Cy3-, and Cy5-conjugated secondary antibodies were used (Jackson Immunoresearch).

transgenic mouse line developed (HBG3)

Facs analysis done on HBG3 mouse line showed 95% motor neurons.

Transplantation: motor neuron axon development studied in chick embryos with rodamine conjugated bungarotoxin or other relevent antibodies.

FUTURE DIRECTION:
Future direction will be to test the ES derived motor neurons in vivo for activity. If they can turn impulses into action then the possibility of using them therpeutically would seem that bit closer