Format

Send to

Choose Destination
Sci Rep. 2015 Jun 11;5:11319. doi: 10.1038/srep11319.

A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture.

Author information

1
Center for Molecular Medicine, National Heart, Lung and Blood Institute, Bethesda, Maryland.
2
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland.
3
Center for Regenerative Medicine, National Institutes of Health, Bethesda, Maryland.
4
1] Center for Molecular Medicine, National Heart, Lung and Blood Institute, Bethesda, Maryland [2] Faculty of Health Sciences, University of Macau, China.

Abstract

Factors limiting the adoption of iPSC technology include the cost of developing lines and the time period that it takes to characterize and bank them, particularly when integration free, feeder free, and Xeno-free components are used. In this manuscript we describe our optimization procedure that enables a single technician to make 20-40 lines at a time in a 24-96 well format in a reliable and reproducible fashion. Improvements spanned the entire workflow and included using RNA virus, reducing cytotoxicity of reagents, developing improved transfection and freezing efficiencies, modifying the manual colony picking steps, enhancing passaging efficiency and developing early criteria of success. These modifications allowed us to make more than two hundred well-characterized lines per year.

PMID:
26066579
PMCID:
PMC4464084
DOI:
10.1038/srep11319
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center