Abstract
We report that iPS cells can be safely and effectively generated from fresh human cord blood (CB) cells with Sendai virus (SeV) vector carrying reprogramming factors OCT3/4, SOX2, KLF4, and c-MYC. The SeV vector is a single strand RNA virus having no DNA phase, and selectively infects the freshly isolated CD34+ CD45low+ fraction of CB cells corresponding to hematopoietic progenitors. Approximately twenty ES cell-like colonies emerged from 1 x 104 freshly isolated CD34+ CB cells around 18 days after SeV infection and were selected for passage to reduce the frequency of the remaining SeV-infected cells. The complete elimination of viral constructs was confirmed after several passages by immunostaining with monoclonal antibody against hemagglutinin-neuraminidase (HN) and by RT-PCR analysis. Five ES cell-like clones were selected to examine their in vitro potential for three germ layer differentiation and their capacity for teratoma formation. Generation of non-integrating Sendai virus (SeV) iPS cells from CB cells may be an important step to provide allogeneic iPS cell-derived therapy in the future.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antigens, CD34 / metabolism
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Biomarkers / metabolism
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Cell Differentiation
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Cell Proliferation
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Cell Separation
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Cells, Cultured
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Cellular Reprogramming
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Coculture Techniques
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Feeder Cells
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Fetal Blood / cytology*
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Gene Expression Profiling / methods
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Gene Expression Regulation, Developmental
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Genetic Vectors
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Hematopoietic Stem Cells / immunology
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Hematopoietic Stem Cells / metabolism
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Hematopoietic Stem Cells / physiology*
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Humans
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Induced Pluripotent Stem Cells / immunology
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Induced Pluripotent Stem Cells / metabolism
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Induced Pluripotent Stem Cells / physiology*
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Karyotyping
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Kruppel-Like Factor 4
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Kruppel-Like Transcription Factors / genetics
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Kruppel-Like Transcription Factors / metabolism
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Octamer Transcription Factor-3 / genetics
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Octamer Transcription Factor-3 / metabolism
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Oligonucleotide Array Sequence Analysis
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Polymerase Chain Reaction
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Proto-Oncogene Proteins c-myc / genetics
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Proto-Oncogene Proteins c-myc / metabolism
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SOXB1 Transcription Factors / genetics
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SOXB1 Transcription Factors / metabolism
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Sendai virus / genetics
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Time Factors
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Transfection
Substances
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Antigens, CD34
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Biomarkers
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KLF4 protein, human
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Kruppel-Like Factor 4
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Kruppel-Like Transcription Factors
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Octamer Transcription Factor-3
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Proto-Oncogene Proteins c-myc
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SOXB1 Transcription Factors