A major focus of our work is to generate of transplantable haematopoietic stem cells (HSCs) from human pluripotent stem cells for tissue repair and regeneration, disease modeling and the testing of pharmaceuticals and other therapeutic products.
Hematopoietic stem cells generated from patient-derived iPSCs may offer a great alternative for patients in need of bone marrow transplants when a matching donor cannot be found. However, successful bone marrow engraftment using in vitro generated iHSCs has been inconsistent in animal models, indicating heterogeneity in the transplanted cells. The transcription factor hepatic leukemia factor (HLF) is one of most specific HSC marker genes identified, with highly selective expression in HSC-enriched subpopulations of cord blood-derived HSCs, peripheral blood, adult bone marrow and the foetal liver. Furthermore, cord blood derived HLF-expressing cells encompass all stem cell activity in vivo during serial transplantation in mice. We are characterising HLF expression during iHSC generation using an HLF reporter cell line, explore the transcriptional profiles and engraftment potential of HLF-expressing cells and assess genes regulated by HLF using Cut-and-Run techniques to identify DNA binding sites for HLF. New insights gained from this project will aid in the identification of engrafting cell populations facilitating the predictability of transplantation outcomes.
We have generated a hlf reporter cell line and characterised hlf expression during HSC development from human iPSCs in vitro. This led to the identification of several subpopulations which we will further investigate in transcriptomic assays and test their transplantablity in vivo.
Expression of the hlf transcription factor hlf coincides with the formation of hematogenic endothelium in embryonic bodies generated from human iPSCs.
Dr Jana Hagen; Ritika Saxena; Elefanty lab MCRI