Protocol detail

Differentiation of iPSC-derived Motoneurons

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The differentiation of motoneurons derived from iPSCs was performed as previously described [21 (link)]. Briefly, iPSCs were dissociated with dispase (Gibco, 1 mg/mL) or accutase (Stem cell) and cultured in MN induction medium, including Neurobasal medium (Invitrogen), DMEM/F12 (Hyclone) at 1:1, 0.5 × B27 supplement (50×) (Gibco), 0.5 × N2 supplement (100×) (Gibco), and 1 × GlutaMAX (100×) (Gibco). Different combinations of DMH1 (Selleck Chemicals, Houston, TX, USA), retinoic acid (RA) (Sigma-Aldrich), CHIR99021 (Selleck), SB431542 (Selleck), valproic acid (Sigma-Aldrich), purmorphamine (Pur) (Selleck), and DAPT (Selleck) were added to the medium at different stages (Figure 2A).

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Zhang Y., Zeng B., Gu A., Kang Q., Zhao M., Peng G., Zhou M., Liu W., Liu M., Ding L., Liang D., Liu X, & Liu M. (2022). Damaged DNA Is an Early Event of Neurodegeneration in Induced Pluripotent Stem Cell-Derived Motoneurons with UBQLN2P497H Mutation. International Journal of Molecular Sciences, 23(19), 11333.

Publication 2022

dispase accutase Neurobasal medium B27 supplement (50×) N2 supplement GlutaMAX retinoic acid (RA) CHIR99021 SB431542 valproic acid

Accutase Chir99021 Dapt Dispase Ipscs Motoneurons Purmorphamine Retinoic acid Sb431542 Stem cell Supplement Valproic acid

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Variable analysis

independent variables

DMH1 (Selleck Chemicals, Houston, TX, USA)
Retinoic acid (RA) (Sigma-Aldrich)
CHIR99021 (Selleck)
SB431542 (Selleck)
Valproic acid (Sigma-Aldrich)
Purmorphamine (Pur) (Selleck)
DAPT (Selleck)

dependent variables

Differentiation of motoneurons derived from iPSCs

control variables

Neurobasal medium (Invitrogen)
DMEM/F12 (Hyclone) at 1:1
0.5 × B27 supplement (50×) (Gibco)
0.5 × N2 supplement (100×) (Gibco)
1 × GlutaMAX (100×) (Gibco)

controls

Not explicitly mentioned
Not explicitly mentioned

Annotations

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