Generation of Human Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells Using Sendai Virus

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1357; p. 23
• Main Authors: Soares, Filipa A C, Pedersen, Roger A, Vallier, Ludovic
• Format: Journal Article
• Language: English
• Published: United States 01.01.2016
• Subjects: 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; Human induced pluripotent stem cells; Peripheral blood mononuclear cells; Reprogramming; Sendai virus; Erythroblast

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...