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Generation of hematopoietic stem cells (HSCs) from pluripotent stem cells (PSCs) could potentially provide unlimited HSCs for clinical transplantation, a curative treatment for numerous blood diseases. However, to date, bona fide HSC generation has been largely unsuccessful in vitro. We have previously described proof of concept for in vivo HSC generation from PSCs via teratoma formation. However, our first-generation system was complex and the output low. Here, we further optimize this technology and demonstrate the following: (1) simplified HSC generation using transcription factor overexpression; (2) improved HSC output using c-Kit-deficient host mice, and (3) that teratomas can be transplanted and cryopreserved. We demonstrate that overexpression of Gfi1b, c-Fos, and Gata2, previously reported to transdifferentiate fibroblasts into hematopoietic progenitors in vitro, can induce long-term HSC formation in vivo. Our in vivo system provides a useful platform to investigate new strategies and re-evaluate existing strategies to generate HSCs and study HSC development.

Original publication




Journal article


Stem Cell Reports

Publication Date





1024 - 1033


Gata2, Gfi1b, cFos, hematopoietic stem cell, hemogenic endothelium, induced pluripotent stem cell, teratomas, Animals, Biomarkers, Cell Transdifferentiation, Cellular Reprogramming, Endothelial Cells, Fibroblasts, GATA2 Transcription Factor, Gene Expression, Gene Order, Genes, fos, Genetic Vectors, Hematopoietic Stem Cells, Immunophenotyping, Induced Pluripotent Stem Cells, Mice, Proto-Oncogene Proteins, Proto-Oncogene Proteins c-kit, Repressor Proteins, Stem Cell Transplantation, Teratoma