Rats were used for transplantation of cultured neurons obtained from K048 human neural stem cells (hNSCs).
Immunocytochemistry was performed on rat brains or cultured neuronal cells with monoclonal mouse anti-Class III-tubulin (TuJ1), 1:4000 (Covance); mouse anti-IsletI 1:50 (Developmental Studies Hybridoma Bank, Iowa); goat anti-CHat, 1:100; polyclonal rabbit anti-synapsin I, 1:500; mouse anti-NeuN, 1:100; mouse anti-human nuclei, 1:20; polyclonal rabbit anti-cytochrome P450scc, 1:400; rabbit anti-TH, 1:500; mouse anti-GalC, 1:100; polyclonal rabbit anti-GFAP; 1:1000 (all from Chemicon), rabbit anti-GABA, 1:1000 (Sigma), rabbit anti-glutamate, 1:5000 (Sigma), or mouse anti-human Nestin, 1:200 (gift of C.A. Messam, NIH). Secondary antibodies conjugated to Alexa Fluo 594 were use for fluorescent detection. Cell nuclei were DAPI stained (Sigma).
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Cholinergic Neurons from Stem Cells—Another Step Toward a Distant Goal
