Alzheimer’s disease

Diseased iPSC lines - Alzheimer's disease

The iPSC lines ICGi008-A and ICGi008-B (https://hpscreg.eu/cell-line/ICGi008-A, https://hpscreg.eu/cell-line/ICGi008-B) were generated from dermal fibroblasts using episomal vectors expressing pluripotency factors. Dermal fibroblasts were obtained from a 55 year old male Сaucasian familial Alzheimer’s disease (AD) patient carrying heterozygous V717I mutation in the APP gene. The generated iPSC lines maintained the original APP genotype, expressed pluripotency markers, exhibited a normal karyotype and retained the ability to differentiate into cell types of the three germ layers. The iPSC lines will be useful for the study of the AD molecular and cellular mechanisms and drug screening.

Diseased iPSC lines - Alzheimer's disease
Characterization of ICGi008-A and ICGi008-B iPSC lines. A. Morphology of the iPSC colonies. B. Immunofluorescence staining for pluripotency markers NANOG, OCT3/4, SOX2, TRA-1-60. C. Analysis of pluripotency markers (OCT4, NANOG, SOX2) expression by RT-qPCR. D. Karyotypes (G-banding) (46, XX) of ICGi008-A and ICGi008-B iPSC lines. E. Mutation analysis. Restriction fragment length polymorphism analysis of exon 17 of the APP gene in ICGi008-A and ICGi008-B iPSC lines, patient’s fibroblasts (m55Alz) and HEK293FT (control). Products of hydrolysis by Ksp22I (120 bp and 199 bp) correspond to mutant allele, 319 bp product corresponds to wild type allele. F. Mutation analysis. Sanger sequencing of exon 17 of the APP gene in HEK293FT (control), patient’s fibroblasts (m55Alz) and iPSC lines (ICGi008-A and ICGi008-B) showing a G>A substitution in one of the alleles (black arrows). G. In vitro differentiation. Immunofluorescence staining for differentiation markers αSMA (mesoderm), NF200 (ectoderm), CK18 and GATA6 (endoderm) after spontaneous differentiation of ICGi008-A and ICGi008-B iPSC lines in ebmryoid bodies. The scale bar A - 200 μm, B, G - 100 μm.

The iPSC lines ICGi008-A and ICGi008-B (https://hpscreg.eu/cell-line/ICGi008-A, https://hpscreg.eu/cell-line/ICGi008-B) were generated from dermal fibroblasts using episomal vectors expressing pluripotency factors. Dermal fibroblasts were obtained from a 55 year old male Сaucasian familial Alzheimer’s disease (AD) patient carrying heterozygous V717I mutation in the APP gene. The generated iPSC lines maintained the original APP genotype, expressed pluripotency markers, exhibited a normal karyotype and retained the ability to differentiate into cell types of the three germ layers. The iPSC lines will be useful for the study of the AD molecular and cellular mechanisms and drug screening.