Fig. 1

Establishment of human embryonic stem cell (hESC) lines with mCherry or familial Alzheimer’s disease (fAD) mutations. (a) Schematic diagram showing the procedures for the establishment of the Alzheimer’s disease (AD) human pluripotent stem cell (hPSC) lines and the generation of AD hPSC-derived AD cerebral organoids (COs) and AD neurons. hPSCs were stably transfected with lentiviral vectors containing mCherry or AD genes with fAD mutations. Dissociated single cells were obtained and plated onto the feeder layer. After expansion, single cell-derived colonies were manually selected, and each hPSC line was established by separate expansion. AD COs or AD neurons were generated from each hPSC line. CAG; CMV early enhancer/chicken β-actin. (b) The levels of amyloid precursor protein (APP) and Presenilin1 (PSEN1) expression in the CmC, CA, or CAP hESC lines were determined by western blot analysis. Red squares represent the selected CA or CAP hESC lines based on their APP and/or PSEN1 expression levels. (c) Table summarizing the control and AD hESC lines established in this study. (d) Representative fluorescence microscope images of the CmC, CA, and CAP hESCs. Scale bar = 200 μm. (e) Images showing the alkaline phosphatase activity in the CmC, CA, and CAP hESCs. (f) Representative immunofluorescence images representing the expression of Oct4, Nanog, SSEA4, and Tra-1-60 in the CmC, CA, and CAP hESCs. Scale bar = 100 μm