Rapid and efficient generation of transgene-free iPSC from a small volume of cryopreserved blood

Zhou, H

Martinez, H

Sun, B

Li, A

Zimmer, M

Katsanis, N

Davis, EE

Kurtzberg, J

Lipnick, S

Noggle, S

Rao, M

Chang, S

2022-03-23T21:02:39Z

2022-03-23T21:02:39Z

2015

2022-03-23T21:02:38Z

© The Author(s) 2015.Human peripheral blood and umbilical cord blood represent attractive sources of cells for reprogramming to induced pluripotent stem cells (iPSCs). However, to date, most of the blood-derived iPSCs were generated using either integrating methods or starting from T-lymphocytes that have genomic rearrangements thus bearing uncertain consequences when using iPSC-derived lineages for disease modeling and cell therapies. Recently, both peripheral blood and cord blood cells have been reprogrammed into transgene-free iPSC using the Sendai viral vector. Here we demonstrate that peripheral blood can be utilized formedium-throughput iPSC production without the need to maintain cell culture prior to reprogramming induction. Cell reprogramming can also be accomplished with as little as 3000 previously cryopreserved cord blood cells under feeder-free and chemically defined Xeno-free conditions that are compliant with standard Good Manufacturing Practice (GMP) regulations. The first iPSC colonies appear 2–3 weeks faster in comparison to previous reports. Notably, these peripheral blood- and cord bloodderived iPSCs are free of detectable immunoglobulin heavy chain (IGH) and T cell receptor (TCR) gene rearrangements, suggesting they did not originate from B- or T- lymphoid cells. The iPSCs are pluripotent as evaluated by the scorecard assay and in vitro multi lineage functional cell differentiation. Our data show that small volumes of cryopreserved peripheral blood or cord blood cells can be reprogrammed efficiently at a convenient, cost effective and scalable way. In summary, our method expands the reprogramming potential of limited or archived samples either stored at blood banks or obtained from pediatric populations that cannot easily provide large quantities of peripheral blood or a skin biopsy.

1550-8943

https://hdl.handle.net/10161/24702

Stem Cell Reviews and Reports

10.1007/s12015-015-9586-8

Rapid and efficient generation of transgene-free iPSC from a small volume of cryopreserved blood

Journal article

Davis, EE|0000-0002-2412-8397

Kurtzberg, J|0000-0002-3370-0703

652

665

4

Duke

School of Medicine

Faculty

Basic Science Departments

Clinical Science Departments

Institutes and Centers

Cell Biology

Pathology

Pediatrics

Duke Cancer Institute

Institutes and Provost's Academic Units

Initiatives

Duke Innovation & Entrepreneurship

Pediatrics, Transplant and Cellular Therapy

11