Human erythroid progenitor cell line comprising HPV E6/E7 operably linked to an inducible promoter and method for producing human enucleated red blood cells

摘要

Provided are: a method for producing an immortalized human erythroid progenitor cell line, enabling efficient and stable production of enucleated red blood cells; and a method for producing human enucleated red blood cells from a human erythroid progenitor cell line obtained by the aforementioned production method. An expression cassette capable of inducing expression of HPV-E6/E7 genes in the presence of DOX was introduced into the genomic DNA of blood stem cells. Then, the blood stem cells were cultured in the presence of DOX and a blood growth factor. Thereby, immortalized cell lines of human erythroid progenitor cells were established. Further, it was revealed that culturing the cell lines under a condition where the expression of the HPV-E6/E7 genes was not induced enabled differentiation induction into enucleated red blood cells at a high ratio.

主权项

1. A method for producing a human erythroid progenitor cell line for producing human enucleated red blood cells, the method comprising the steps of:
introducing into human hematopoietic stem cells an expression cassette comprising E6 and E7 genes of human papillomavirus type 16 operably linked to a promoter responsive to an added external stimulus; culturing the human hematopoietic stem cells having the expression cassette introduced therein in the presence of the external stimulus and at least one blood growth factor selected from the group consisting of SCF, EPO, TPO and DEX under conditions to produce human erythroid progenitor cells, and identifying human erythroid progenitor cells in the culture.