Generation of Human Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells Using Sendai Virus
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 exp...
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| Published in | Methods in molecular biology (Clifton, N.J.) Vol. 1357; p. 23 |
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| Main Authors | , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
01.01.2016
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| Subjects | |
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| 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. |
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| AbstractList | 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. |
| Author | Soares, Filipa A C Pedersen, Roger A Vallier, Ludovic |
| Author_xml | – sequence: 1 givenname: Filipa A C surname: Soares fullname: Soares, Filipa A C email: facs2@cam.ac.uk organization: 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 – sequence: 2 givenname: Roger A surname: Pedersen fullname: Pedersen, Roger A organization: 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 – sequence: 3 givenname: Ludovic surname: Vallier fullname: Vallier, Ludovic organization: Wellcome Trust Sanger Institute, Hinxton, UK |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/25687300$$D View this record in MEDLINE/PubMed |
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| Keywords | Human induced pluripotent stem cells Peripheral blood mononuclear cells Reprogramming Sendai virus Erythroblast |
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| Snippet | This protocol describes the efficient isolation of peripheral blood mononuclear cells from circulating blood via density gradient centrifugation and subsequent... |
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| SubjectTerms | Animals Cell Culture Techniques - methods Cell Separation - methods Cells, Cultured Cellular Reprogramming Cellular Reprogramming Techniques - methods Culture Media Cytokines - pharmacology Erythroblasts - cytology Erythroblasts - drug effects Erythroblasts - virology Fibroblasts - cytology Genetic Vectors - genetics Humans Induced Pluripotent Stem Cells - cytology Kruppel-Like Factor 4 Kruppel-Like Transcription Factors - genetics Kruppel-Like Transcription Factors - physiology Leukocytes, Mononuclear - cytology Leukocytes, Mononuclear - drug effects Leukocytes, Mononuclear - virology Mice Octamer Transcription Factor-3 - genetics Octamer Transcription Factor-3 - physiology Proto-Oncogene Proteins c-myc - genetics Proto-Oncogene Proteins c-myc - physiology Recombinant Proteins - genetics Sendai virus - genetics SOXB1 Transcription Factors - genetics SOXB1 Transcription Factors - physiology |
| Title | Generation of Human Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells Using Sendai Virus |
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