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Activation of KLF1 enhances the differentiation and maturation of red blood cells from human pluripotent stem cells

Research output: Contribution to journalArticle

Original languageEnglish
Pages (from-to)886–897
Number of pages12
JournalStem Cells
Volume35
Issue number4
Early online date19 Jan 2017
DOIs
DateAccepted/In press - 8 Dec 2016
DateE-pub ahead of print - 19 Jan 2017
DatePublished (current) - Apr 2017

Abstract

Blood transfusion is widely used in the clinic but the source of red blood cells (RBCs) is dependent on donors, procedures are susceptible to transfusion-transmitted infections and complications can arise from immunological incompatibility. Clinically-compatible and scalable protocols that allow the
production of RBCs from human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) have been described but progress to translation has been hampered by poor maturation and fragility of the resultant cells. Genetic programming using transcription factors has been used to drive lineage determination and differentiation so we used this approach to assess whether exogenous expression of the Erythroid Krüppel-like factor 1 (EKLF/KLF1) could augment the differentiation and stability of iPSC-derived RBCs. To activate KLF1 at defined time-points during later stages of the differentiation process and to avoid transgene silencing that is commonly observed in differentiating PSCs, we targeted a tamoxifen-inducible KLF1-ERT2 expression cassette into the AAVS1 locus. Activation of KLF1 at day 10 of the differentiation process when hematopoietic progenitor cells were present, enhanced erythroid commitment and differentiation. Continued culture resulted the appearance of more enucleated cells when KLF1 was activated which is possibly due to their more robust morphology. Globin profiling indicated that these conditions produced embryonic-like erythroid cells. This study demonstrates the successful use of an inducible genetic programing strategy that could be applied to the production of many other cell lineages from hiPSCs with the integration of programming factors into the AAVS1 locus providing a safer and more reproducible route to the clinic.

    Research areas

  • Erythroid differentiation, Induced pluripotent stem cells, Transcription factors, Gene delivery systems in vivo or in vitro

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    Rights statement: This is the final published version of the article (version of record). It first appeared online via Wiley at http://onlinelibrary.wiley.com/doi/10.1002/stem.2562/abstract. Please refer to any applicable terms of use of the publisher.

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    Rights statement: This is the final published version of the article (version of record). It first appeared online via Wiley at http://onlinelibrary.wiley.com/doi/10.1002/stem.2562/abstract. Please refer to any applicable terms of use of the publisher.

    Final published version, 960 KB, PDF document

    Licence: CC BY

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