title: Reliable generation of induced pluripotent stem cells from human lymphoblastoid cell lines.
abstract: Patient-specific induced pluripotent stem cells (iPSCs) hold great promise for many applications, including disease modeling to elucidate mechanisms involved in disease pathogenesis, drug screening, and ultimately regenerative medicine therapies. A frequently used starting source of cells for reprogramming has been dermal fibroblasts isolated from skin biopsies. However, numerous repositories containing lymphoblastoid cell lines (LCLs) generated from a wide array of patients also exist in abundance. To date, this rich bioresource has been severely underused for iPSC generation. We first attempted to create iPSCs from LCLs using two existing methods but were unsuccessful. Here we report a new and more reliable method for LCL reprogramming using episomal plasmids expressing pluripotency factors and p53 shRNA in combination with small molecules. The LCL-derived iPSCs (LCL-iPSCs) exhibited identical characteristics to fibroblast-derived iPSCs (fib-iPSCs), wherein they retained their genotype, exhibited a normal pluripotency profile, and readily differentiated into all three germ-layer cell types. As expected, they also maintained rearrangement of the heavy chain immunoglobulin locus. Importantly, we also show efficient iPSC generation from LCLs of patients with spinal muscular atrophy and inflammatory bowel disease. These LCL-iPSCs retained the disease mutation and could differentiate into neurons, spinal motor neurons, and intestinal organoids, all of which were virtually indistinguishable from differentiated cells derived from fib-iPSCs. This method for reliably deriving iPSCs from patient LCLs paves the way for using invaluable worldwide LCL repositories to generate new human iPSC lines, thus providing an enormous bioresource for disease modeling, drug discovery, and regenerative medicine applications.
authors.affiliation: Regenerative Medicine Institute, F. Widjaja Foundation Inflammatory Bowel and Immunobiology Research Institute, and Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Barrett
authors.affiliation: Regenerative Medicine Institute, F. Widjaja Foundation Inflammatory Bowel and Immunobiology Research Institute, and Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, USA.
authors.forename: Loren
authors.initials: L
authors.lastname: Ornelas
authors.affiliation: Regenerative Medicine Institute, F. Widjaja Foundation Inflammatory Bowel and Immunobiology Research Institute, and Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, USA.
authors.forename: Nicole
authors.initials: N
authors.lastname: Yeager
authors.affiliation: Regenerative Medicine Institute, F. Widjaja Foundation Inflammatory Bowel and Immunobiology Research Institute, and Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, USA.
authors.forename: Berhan
authors.initials: B
authors.lastname: Mandefro
authors.affiliation: Regenerative Medicine Institute, F. Widjaja Foundation Inflammatory Bowel and Immunobiology Research Institute, and Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, USA.
authors.forename: Anais
authors.initials: A
authors.lastname: Sahabian
authors.affiliation: Regenerative Medicine Institute, F. Widjaja Foundation Inflammatory Bowel and Immunobiology Research Institute, and Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, USA.
authors.forename: Lindsay
authors.initials: L
authors.lastname: Lenaeus
authors.affiliation: Regenerative Medicine Institute, F. Widjaja Foundation Inflammatory Bowel and Immunobiology Research Institute, and Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, USA.
authors.forename: Stephan R
authors.initials: SR
authors.lastname: Targan
authors.affiliation: Regenerative Medicine Institute, F. Widjaja Foundation Inflammatory Bowel and Immunobiology Research Institute, and Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, USA.
authors.forename: Clive N
authors.initials: CN
authors.lastname: Svendsen
authors.affiliation: Regenerative Medicine Institute, F. Widjaja Foundation Inflammatory Bowel and Immunobiology Research Institute, and Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, USA dhruv.sareen@cshs.org.
authors.forename: Dhruv
authors.initials: D
authors.lastname: Sareen
mesh_terms: D002460:Cell Line; D065150:Cellular Reprogramming; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D008214:Lymphocytes / Q000166:cytology* / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000235:genetics
chemical_list: C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53
keywords: B lymphocytes; Developmental biology; Endoderm; Induced pluripotent stem cells; Neural differentiation; Neuron; Pluripotent stem cells; Reprogramming