Method for preparing hemogenic endothelium cell and method for preparing hematopoietic stem cell or hematopoietic stem and progenitor cell

A method for preparing hematopoietic endothelial cells, comprising:

• (a) providing hematopoietic mesodermal cells comprising exogenously introduced nucleic acid sequences encoding transcription factors LCOR, HOXA9, HOXAS, RUNX1, ERG and an exogenously introduced nucleic acid sequence encoding reverse tetracycline transcriptional activator (rtTA) or a cell culture comprising the sequence modified hematopoietic mesodermal cells;
• wherein the nucleic acid sequences encoding the transcription factors LCOR, HOXA9, HOXAS, RUNX1, ERG and the nucleic acid sequence encoding rtTA are integrated in the genome of the hematopoietic mesodermal cells:
• the nucleic acid sequences encoding the transcription factors LCOR, HOXA9, HOXAS, RUNX1, ERG and the nucleic acid sequence encoding rtTA are operably linked to an inducible promoter, wherein the inducible promoter is a tetracycline-inducible promoter:
• the exogenous introduction of the nucleic acid sequences is carried out by a lentiviral vector:
• (b) culturing the hematopoietic mesodermal cells or a cell culture comprising the sequence modified hematopoietic mesodermal cells in a hematopoietic endothelial differentiation medium;
• and
• (c) inducing the hematopoietic mesodermal cells to express the transcription factors LCOR, HOXA9, HOXAS, RUNX1, and ERG by adding tetracycline or doxycycline to the hematopoietic endothelial differentiation medium;
• wherein step (c) is performed on days 1-4 or days 4-7 of the culturing in step (b).