title: Generation of Human Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells Using Sendai Virus.
abstract: This protocol describes the efficient isolation of peripheral blood mononuclear cells from circulating blood via density gradient centrifugation and subsequent generation of integration-free human induced pluripotent stem cells. Peripheral blood mononuclear cells are cultured for 9 days to allow expansion of the erythroblast population. The erythroblasts are then used to derive human induced pluripotent stem cells using Sendai viral vectors, each expressing one of the four reprogramming factors Oct4, Sox2, Klf4, and c-Myc.
authors: Anne McLaren Laboratory for Regenerative Medicine and Department of Surgery, Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, West Forvie Building, Robison Way, CB2 0SZ, Cambridge, UK. facs2@cam.ac.uk.
authors: Filipa A C
authors: FA
authors: Soares
authors: Anne McLaren Laboratory for Regenerative Medicine and Department of Surgery, Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, West Forvie Building, Robison Way, CB2 0SZ, Cambridge, UK.
authors: Roger A
authors: RA
authors: Pedersen
authors: Anne McLaren Laboratory for Regenerative Medicine and Department of Surgery, Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, West Forvie Building, Robison Way, CB2 0SZ, Cambridge, UK.|Wellcome Trust Sanger Institute, Hinxton, UK.
authors: Ludovic
authors: L
authors: Vallier
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D000067470:Cellular Reprogramming Techniques / Q000379:methods*; D003470:Culture Media; D016207:Cytokines / Q000494:pharmacology; D004900:Erythroblasts / Q000166:cytology / Q000187:drug effects / Q000821:virology; D005347:Fibroblasts / Q000166:cytology; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000502:physiology; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000187:drug effects / Q000821:virology; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000502:physiology; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000502:physiology; D029082:Sendai virus / Q000235:genetics*
chemical_list: D003470:Culture Media; D016207:Cytokines; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C489427:MYC protein, human; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D016271:Proto-Oncogene Proteins c-myc; D011994:Recombinant Proteins; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors
keywords: Erythroblast; Human induced pluripotent stem cells; Peripheral blood mononuclear cells; Reprogramming; Sendai virus