L glutamine

Two common marmoset ES cell lines, No. 40 (female, 46XX) and DSY127 (male, 46XY) were used in the current study. The No. 40 line was established previously^{44 (link)}, and DSY127 was kindly provided by Sumitomo Dainippon Pharma Co., Ltd. (Tokyo, Japan). These cells were cultured as described previously^{45 (link)}. In brief, cjESCs were cultured on 30 Gy-irradiated mouse embryonic fibroblasts (MEFs) in ES medium (ESM) consisting of 1x KnockOut DMEM (Thermo Fisher) supplemented with 20% KnockOut Serum Replacement (Thermo Fisher), 1mM L-glutamine (Nakalai Tesque), 1% non-essential amino acids (Sigma), 0.2 mM 2-mercaptoethanol (Sigma) and 10 ng/ml fibroblast growth factor 2 (Peprotech). The KI cjESC lines generated in the current study will be distributed by the corresponding authors upon request.

The H-1 mESC line was originally isolated from C3H/He mice [23 (link)]. mESCs were maintained in Dulbecco’s modified Eagle’s medium (4.5 g/l glucose) with L-glutamine, without sodium pyruvate, (Nacalai Tesque, Japan) supplemented with 15% foetal bovine serum (Gibco, USA), 1000 U/ml Stem Sure Leukemia Inhibitory Factor (mouse recombinant solution; Wako), 0.1 mM Stem Sure 2-mercaptoethanol solution (Wako), and penicillin–streptomycin (Gibco). Cells were grown on mitomycin C (Kyowa Kirin, Japan)-treated mouse embryonic fibroblast feeder cells (C57BL/6J) at 37°C in a humidified incubator with 5% CO₂. For chemical stress treatments, mESCs were cultured in ESGRO Complete Plus serum-free clonal grade medium (Merck Millipore, Germany) on gelatine (Sigma, USA)-coated dishes without feeder cells.

For myogenic differentiation, we modified a previously reported protocol (Fig. 2a). Briefly, iPSC^MyoD were trypsinized, dissociated to single cells, and seeded on Matrigel (BD Biosciences, San Diego, CA, USA)-coated plates at a density from 4 × 10³ to 5 × 10⁴ cells/cm² (cell line-specific) in 20% (v/v) knockout serum replacement (KSR) human iPSC medium with 10 µM Y-27632. After 24 h, Dox (LKT Laboratories, St. Paul, MN, USA) was added at 1 µg/mL. On day 2, the medium was replaced with high glucose (4.5 g/L) Dulbecco’s modified Eagle’s medium (DMEM) (Nacalai Tesque) containing 5% (v/v) KSR (Thermo Fisher Scientific), 1 µg/mL Dox, 1 mM L-glutamine (Thermo Fisher Scientific), and 0.1 mM 2-mercaptoethanol (2-ME) (Thermo Fisher Scientific). The medium was changed daily. For the transient glucose deprivation experiment, the medium was replaced with glucose-free DMEM/Ham’s F-12 (Nacalai Tesque) containing 1 µg/mL Dox, 1 mM L-glutamine and 0.1 mM 2-ME for 24 h prior to the glycogen analysis. For the rhGAA rescue experiment, 1 µM Myozyme (rhGAA) (Sanofi Genzyme, Cambridge, MA, USA) was added to the medium for the last 3 days unless otherwise specified.

LNCaP cells, MCF7 cells, MDA-MB-231 cells, Caco-2 cells, HEK293 cells, and Cos7 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Nacalai Tesque, Kyoto, Japan); PC3 cells were cultured in RPMI 1640 medium (Nacalai Tesque); and A375 cells were cultured in Eagle’s Minimum Essential Medium (Wako Pure Chemical Industries, Osaka, Japan). All media were supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, Saint Louis, MO, USA), 20 mM l-glutamine (Nacalai Tesque) and 100 µ/mL penicillin–streptomycin (Nacalai Tesque). PrS cells were purchased from Lonza (Cat. No. CC-2508; Basel, Switzerland) and cultured in Stromal cell basal medium supplemented with growth factors (Lonza). Male BALB/c nude mice (7-weeks-old) were purchased from Japan SLC (Shizuoka, Japan). The animal experiments conducted in this study were approved by the Shiga University of Medical Science Animal Care and Use Committee according to the Animal Research Reporting of In Vivo Experiments guidelines..

Human neuroblastoma SH-SY5Y cells were obtained from the Institute of Bioscience (IBS), Universiti Putra Malaysia (UPM). SH-SY5Y cells are established as an NDDs model and could be differentiated into terminal neurons by retinoic acid (RA)-treatment. The cells were maintained in a 1:1 mixture of Dulbecco Modified Eagle Medium and Ham’s F12 (DMEM/Hams’ F12) (Nacalai, Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS), 2 mM of L-glutamine and 1% penicillin/streptomycin (Nacalai, Kyoto, Japan) in a 5% CO₂ incubator at 37 °C. Cell culture media were replaced every 2 days, and, at 80–90% of confluence, the cells were sub-cultured [25 (link)].