RRC ID 55771
Author Nishishita N, Takenaka C, Fusaki N, Kawamata S.
Title Generation of human induced pluripotent stem cells from cord blood cells.
Journal J Stem Cells
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.
Volume 6(3)
Pages 101-8
Published 2011
PMID 23264996
MeSH Antigens, CD34 / metabolism Biomarkers / metabolism Cell Differentiation Cell Proliferation Cell Separation Cells, Cultured Cellular Reprogramming Coculture Techniques Feeder Cells Fetal Blood / cytology* Gene Expression Profiling / methods Gene Expression Regulation, Developmental Genetic Vectors Hematopoietic Stem Cells / immunology Hematopoietic Stem Cells / metabolism Hematopoietic Stem Cells / physiology* Humans Induced Pluripotent Stem Cells / immunology Induced Pluripotent Stem Cells / metabolism Induced Pluripotent Stem Cells / physiology* Karyotyping Kruppel-Like Transcription Factors / genetics Kruppel-Like Transcription Factors / metabolism Octamer Transcription Factor-3 / genetics Octamer Transcription Factor-3 / metabolism Oligonucleotide Array Sequence Analysis Polymerase Chain Reaction Proto-Oncogene Proteins c-myc / genetics Proto-Oncogene Proteins c-myc / metabolism SOXB1 Transcription Factors / genetics SOXB1 Transcription Factors / metabolism Sendai virus / genetics Time Factors Transfection
Resource
Cord blood stem cells for research