Three separate human iPSC lines were used in these studies to ensure that the
successful generation of microglia could be reproduced using multiple iPSCs.
Human iPSC line NCRM-1 was obtained from the National Institutes of Health Stem
Cell Resource (Bethesda, MD, USA). Human iPSC lines CS52iALS-C9n6.ISOxx
(referred to as “CS52” hereafter for sake of brevity) and CS29iALS-C9n1.ISOxx
(“CS29” hereafter), corresponding to corrected, isogenic lines derived from two
distinct parental lines generated from ALS patients with
C9orf72 gene mutations, were obtained from Cedars-Sinai
(Los Angeles, CA, USA) (Thiry et al., 2022 (link)). Undifferentiated state of human iPSCs was
assessed by testing for expression of the stem cell markers NANOG and OCT4 using
rabbit anti-NANOG (1/1,000; Abcam; Cambridge, UK; Cat. No. ab21624) and rabbit
anti-OCT4 (1 µg/ml; Abcam; Cat. No. ab19857) or goat anti-OCT3/4 (1/500; Santa
Cruz Biotechnology; Dallas, TX, USA, Cat. No. sc-8628) antibodies, as described
(Chen et al.,
2021 ).
successful generation of microglia could be reproduced using multiple iPSCs.
Human iPSC line NCRM-1 was obtained from the National Institutes of Health Stem
Cell Resource (Bethesda, MD, USA). Human iPSC lines CS52iALS-C9n6.ISOxx
(referred to as “CS52” hereafter for sake of brevity) and CS29iALS-C9n1.ISOxx
(“CS29” hereafter), corresponding to corrected, isogenic lines derived from two
distinct parental lines generated from ALS patients with
C9orf72 gene mutations, were obtained from Cedars-Sinai
(Los Angeles, CA, USA) (Thiry et al., 2022 (link)). Undifferentiated state of human iPSCs was
assessed by testing for expression of the stem cell markers NANOG and OCT4 using
rabbit anti-NANOG (1/1,000; Abcam; Cambridge, UK; Cat. No. ab21624) and rabbit
anti-OCT4 (1 µg/ml; Abcam; Cat. No. ab19857) or goat anti-OCT3/4 (1/500; Santa
Cruz Biotechnology; Dallas, TX, USA, Cat. No. sc-8628) antibodies, as described
(
2021
