title: A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture.
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.
authors.affiliation: Center for Molecular Medicine, National Heart, Lung and Blood Institute, Bethesda, Maryland.
authors.forename: Jeanette
authors.initials: J
authors.lastname: Beers
authors.affiliation: Center for Molecular Medicine, National Heart, Lung and Blood Institute, Bethesda, Maryland.
authors.forename: Kaari L
authors.initials: KL
authors.lastname: Linask
authors.affiliation: Center for Molecular Medicine, National Heart, Lung and Blood Institute, Bethesda, Maryland.
authors.forename: Jane A
authors.initials: JA
authors.lastname: Chen
authors.affiliation: Center for Molecular Medicine, National Heart, Lung and Blood Institute, Bethesda, Maryland.
authors.forename: Lauren I
authors.initials: LI
authors.lastname: Siniscalchi
authors.affiliation: Center for Molecular Medicine, National Heart, Lung and Blood Institute, Bethesda, Maryland.
authors.forename: Yongshun
authors.initials: Y
authors.lastname: Lin
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland.
authors.forename: Wei
authors.initials: W
authors.lastname: Zheng
authors.affiliation: Center for Regenerative Medicine, National Institutes of Health, Bethesda, Maryland.
authors.forename: Mahendra
authors.initials: M
authors.lastname: Rao
authors.affiliation: 1] Center for Molecular Medicine, National Heart, Lung and Blood Institute, Bethesda, Maryland [2] Faculty of Health Sciences, University of Macau, China.
authors.forename: Guokai
authors.initials: G
authors.lastname: Chen
mesh_terms: D018929:Cell Culture Techniques / Q000191:economics / Q000379:methods*; D002460:Cell Line; D065150:Cellular Reprogramming; D000067470:Cellular Reprogramming Techniques / Q000191:economics / Q000379:methods*; D003470:Culture Media / Q000737:chemistry*; D006801:Humans; D057026:Induced Pluripotent Stem Cells
chemical_list: D003470:Culture Media