Multipotent lymphohematopoietic progenitor cells
This invention relates to hematopoietic precursors derived from human embryonic stem cells. In the culture of differentiated cells from human ES cells, the fully committed hematopoietic precursors are CD34+ and CD43+ but not CD45+. If the cells are cultured until they express CD45, then the cells lose the ability to produce differentiated cells of the lymphoid lineages.
1. A purified population of CD34+ CD43+ CD45+ CD41 a− CD235a− cells, wherein the cells are prepared by a method comprising:
(a) preparing a culture of macrophage colony-stimulating factor (M-CSF) deficient stromal cells;
(b) adding human embryonic stem (ES) cells to said culture in medium supplemented with serum;
(c) culturing said ES cells until the culture begins to express CD45;
(d) isolating cells that are both CD34+ cells and CD43+ from the resulting culture; and
(e) separating CD45+ CD41 a− CD235a− cells from the isolated cells of step (d), wherein greater than 95% of the cells separated in (e) are CD34+ CD43+ CD45+ CD41 a− CD235a− cells.
2. A cell culture comprising the purified population of cells of claim 1 .
3. A method for generating a population of CD34+ CD43+ CD45+ cells comprising the steps of:
(a) preparing a culture of macrophage colony-stimulating factor (M-CSF) deficient stromal cells;
(b) adding human ES cells to said culture in medium supplemented with serum;
(c) culturing said ES cells until the culture begins to express CD45;
(d) isolating cells that are both CD34+ and CD43+ cells; and
(e) separating CD45+ CD41 a− CD235a− cells from the isolated cells of step (d), wherein greater than 95% of the cells separated in (f) are CD34+ CD43+ CD45+ CD41 a− CD235a− cells.