A disease-specific iPS cell resource for studying rare and intractable diseases

Abstract

Background: Disease-specific induced pluripotent stem cells (iPSCs) are useful tools for pathological analysis and diagnosis of rare diseases. Given the limited available resources, banking such disease-derived iPSCs and promoting their widespread use would be a promising approach for untangling the mysteries of rare diseases. Herein, we comprehensively established iPSCs from patients with designated intractable diseases in Japan and evaluated their properties to enrich rare disease iPSC resources.

Methods: Patients with designated intractable diseases were recruited for the study and blood samples were collected after written informed consent was obtained from the patients or their guardians. From the obtained samples, iPSCs were established using the episomal method. The established iPSCs were deposited in a cell bank.

Results: We established 1,532 iPSC clones from 259 patients with 139 designated intractable diseases. The efficiency of iPSC establishment did not vary based on age and sex. Most iPSC clones originated from non-T and non-B hematopoietic cells. All iPSC clones expressed key transcription factors, OCT3/4 (range 0.27-1.51; mean 0.79) and NANOG (range 0.15-3.03; mean 1.00), relative to the reference 201B7 iPSC clone.

Conclusions: These newly established iPSCs are readily available to the researchers and can prove to be a useful resource for research on rare intractable diseases.

Keywords: Designated diseases; Rare and intractable diseases; Reprogramming; iPS cells.

Fig. 1

Characteristics of iPSC donors. See also Supplementary Table S2. A Workflow from donor recruitment to depositing iPSCs in Riken Bioresource Bank. B Age distribution of recruited donors

Fig. 2

Establishment of iPSC lines. See also Supplementary Fig. 1. A Flowchart depicting iPSC establishment. B Relationship between sex and reprogramming efficiency (%). C Relationship between sex and doubling time of established iPSC lines. (total n = 1,039; female n = 535; male n = 504). B, C Statistical analysis was performed using Mann–Whitney U test

Fig. 3

Basic characterization of established iPSCs. See also Supplementary Fig. 2. A Representative phase contrast images of iPSC colonies. iPSC clones from the patients with (i) Duchenne muscular dystrophy, (ii) Alexander disease, (iii) Dravet syndrome and (iv) Smith-Magenis syndrome are shown. Scale bars = 100 μm. B Estimation of the cell type from which iPSCs originated; n = 1,532. C Estimation of the number of residual copies of episomal vectors remaining in iPSCs, calculated based on the quantitative values of EBNA and CAG promotors. Dashed lines are drawn at the line corresponding to one copy of remaining vector per cell; n = 1,532. D Correlation between OCT3/4 and NANOG expression in each clone. The single regression equation is plotted on the graph; n = 1,532. E, F Comparison of OCT3/4 (E) and NANOG (F) expression by sex (female n = 798, male n = 734). Values for each clone have been plotted relative to the expression levels in control 201B7 iPSCs. Statistical analysis was performed using the unpaired t-test (E) and Mann–Whitney U test (F)