Generation of human induced pluripotent stem cell-lines by CGaP (WTSI)
Research type
Research Study
Full title
Generation of human induced pluripotent stem cell-lines for research by the Cellular Generation and Phenotyping Facility at the Wellcome Trust Sanger Institute
IRAS ID
148586
Contact name
Chukwuma A. Agu
Contact email
Sponsor organisation
Genome Research Ltd.
Duration of Study in the UK
5 years, 0 months, 0 days
Research summary
Human iPS cells are derived from mature tissue cells whose genetic clocks have been reset to turn them back into stem cells, giving them the ability to develop into almost every cell type in the body. The Cellular Generation and Phenotyping (CGaP) facility will create a research resource of human induced pluripotent stem cells to examine genes in health and disease by deriving iPS cell lines from apparently healthy donors and patients, and examine genetic and epigenetic variation in human genomes and its consequences for gene activity.
Samples (blood, saliva and skin) from patients with inherited DNA repair/replication disorders and patients with a history of strong exposure to DNA damaging agents will be included in this study. These patients were previously recruited and consented for the REC approved study 'Exploring the biological processes underlying mutational signatures identified in patients with inherited disorders and in patients exposed to mutagens' (Dr. Serena Nik-Zainal, REC reference number 13/EE/0302, NRES Committee East of England - Norfolk).
Blood samples only will also be obtained from controls - blood donors from NHS Blood and Transplant, and commercial suppliers.
The principle research objectives are:
- To derive human iPS cells and immortalised cell-lines from apparently healthy people and specific patient groups, and use these cell resources to examine cellular activity to better understand biology in health and disease.- To create human induced pluripotent stem cells, examine genes in health and disease, and examine genetic and epigenetic variation in human genomes ad its consequences for gene activity.
- To perform stability testing on somatic cells in whole blood using anonymised samples from the NHSBT, commercial suppliers and Dr. Nik-Zainal’s study.
The secondary research objective will be to create a research resource of human induced pluripotent stem cells which will be shared with the research community via a third party distributor.
Lay summary of study results: The collection sites of human primary tissue samples and the receiving laboratories, where the human induced pluripotent stem cells (hIPSCs) are derived, are often not on the same site. Thus, the stability of samples prior to derivation constrains the distance between the collection site and the receiving laboratory. To investigate sample stability, we collected blood and held it at room temperature for 5, 24, or 48 hr before isolating peripheral blood mononuclear cells (PBMCs) and reprogramming into IPSCs. Additionally, PBMC samples at 5- and 48-hr time points were frozen in liquid nitrogen for 4 months and reprogrammed into IPSCs. hIPSC lines derived from all time points were pluripotent, displayed no marked difference in chromosomal aberration rates, and differentiated into three germ layers. Reprogramming efficiency at 24- and 48-hr time points was 3- and 10-fold lower, respectively, than at 5 hr; the freeze-thaw process of PBMCs resulted in no obvious change in reprogramming efficiency.
REC name
London - Fulham Research Ethics Committee
REC reference
14/LO/0345
Date of REC Opinion
17 Feb 2014
REC opinion
Favourable Opinion