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Leukemia. 2016 Mar;30(3):674-82. doi: 10.1038/leu.2015.294. Epub 2015 Oct 26.

Reprogramming human B cells into induced pluripotent stem cells and its enhancement by C/EBPα.

Author information

1
Josep Carreras Leukemia Research Institute and School of Medicine, University of Barcelona, Barcelona, Spain.
2
Gene Regulation, Stem Cells and Cancer Program, Centre for Genomic Regulation (CRG) and University Pompeu Fabra (UPF), Barcelona, Spain.
3
Department of Hematology, University Hospital of Salamanca, Instituto Biosanitario de Salamanca (IBSAL), Salamanca, Spain.
4
Servei d'Oncologia Radioteràpica, Hospital Clinic, Barcelona, Spain.
5
Cancer Epigenetics Laboratory, Instituto Universitario de Oncología del Principado de Asturias, Universidad de Oviedo, Asturias, Spain.
6
Hematology Department, Hospital Germans Trias i Pujol, Institut Català d'Oncologia, Badalona, Spain.
7
Differentiation and Cytometry Unit, Hematopoietic Innovative Therapy Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT), Madrid, Spain.
8
Faculty of Medicine, Laboratory of Gene Regulation, University of Tsukuba, Tsukuba, Ibaraki, Japan.
9
Research Center for Stem Cell Engineering and National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.
10
Instituciò Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain.

Abstract

B cells have been shown to be refractory to reprogramming and B-cell-derived induced pluripotent stem cells (iPSC) have only been generated from murine B cells engineered to carry doxycycline-inducible Oct4, Sox2, Klf4 and Myc (OSKM) cassette in every tissue and from EBV/SV40LT-immortalized lymphoblastoid cell lines. Here, we show for the first time that freshly isolated non-cultured human cord blood (CB)- and peripheral blood (PB)-derived CD19+CD20+ B cells can be reprogrammed to iPSCs carrying complete VDJH immunoglobulin (Ig) gene monoclonal rearrangements using non-integrative tetracistronic, but not monocistronic, OSKM-expressing Sendai Virus. Co-expression of C/EBPα with OSKM facilitates iPSC generation from both CB- and PB-derived B cells. We also demonstrate that myeloid cells are much easier to reprogram than B and T lymphocytes. Differentiation potential back into the cell type of their origin of B-cell-, T-cell-, myeloid- and fibroblast-iPSCs is not skewed, suggesting that their differentiation does not seem influenced by 'epigenetic memory'. Our data reflect the actual cell-autonomous reprogramming capacity of human primary B cells because biased reprogramming was avoided by using freshly isolated primary cells, not exposed to cytokine cocktails favoring proliferation, differentiation or survival. The ability to reprogram CB/PB-derived primary human B cells offers an unprecedented opportunity for studying developmental B lymphopoiesis and modeling B-cell malignancies.

PMID:
26500142
DOI:
10.1038/leu.2015.294
[Indexed for MEDLINE]

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