| Literature DB >> 26500142 |
C Bueno1, J L Sardina2, B Di Stefano2, D Romero-Moya1, A Muñoz-López1, L Ariza1, M C Chillón3, A Balanzategui3, J Castaño1, A Herreros4, M F Fraga5, A Fernández5, I Granada1,6, O Quintana-Bustamante7, J C Segovia7, K Nishimura8, M Ohtaka9, M Nakanishi9, T Graf2, P Menendez1,10.
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.Entities:
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Year: 2015 PMID： 26500142 DOI： 10.1038/leu.2015.294
Source DB: PubMed Journal: Leukemia ISSN： 0887-6924 Impact factor: 11.528