The analysis performed in healthy and ALS hNSC spheroids undergoing the rest of differentiation protocols (A’-D) to assess the effects of each differentiation protocol onto the initial steps of the differentiation showed that all protocols were able to induce the formation of HNK1 + cells at TP2 in healthy and ALS hNSC. This evidence concerns the gene B3GAT1 and amyotrophic lateral sclerosis.