Human brain endothelial cells and growth medium and method for expansion of primitive CD34+CD38-bone marrow stem cells

A method of expanding human bone marrow CD34+CD38− hematopoietic progenitor cells, including primitive stem cells, in vitro, comprising the steps of:

• i) isolating and culturing human brain endothelial cells by dissecting segments of blood vessels from the Circle of Willis;
• ii) isolating CD34+CD38− hematopoietic progenitor cells from human bone marrow;
• iii) contacting the isolated CD34+CD38− hematopoietic progenitor cells with the isolated and cultured human brain endothelial cells;
• and
• iv) co-culturing the contacted CD34+CD38− hematopoietic progenitor cells and endothelial cells in the presence of at least one cytokine in an amount sufficient to support aniplification/expanston ofCD34+CD38− hematopoietic progenitor cells.

A method of amnplifgin/expanding human CD34+CD38− hematopoietic progenitor cells, in vitro, which comprises the steps of:

• i) isolating and culturing human brain endothelial cells by dissecting segments ofbood vessels from the Circle of Willis;
• ii) contacting isolated CD34+CD38− hematopoictic progenitor cells with the isolated and cultured human brain endothelial cells;
• and
• iii) co-culturing the contacted CD34+CD38− hematopoietic progenitor cells and endothelial cells in the presence of a mixture of granulocyte-macrophage colony stimulating factor, interleukin-3, stem cell factor and interleukin-6 in an amount sufficient to amplify/expand said CD34+CD38− bematopoietic progenitor cells.

A method of amphifying/expanding human CD34+CD38− hematopoietic progenitor cells, in vitro, which comprises the steps of:

• i) isolating and culturing human brain endothelial cells by dissecting segments of blood vessels from the Circle of Willis;
• ii) isolating the CD34+CD38− hematopoietic progenitor cells from humnan bone marrow;
• iii) contacting the isolated CD34+CD38− hematopoietic progenitor cells with the isolated and cultured humnan brain endothelial cells;
• and
• iv) co-culturg the contacted CD34+CD38− hematopoietic progenitor cells and endothelial cells in the presence of a mixture of granulocyte-macrophage colony stimulating factor, interlekukin-3, stem cell factor and interleukin-6 in an amount sufficient to amplify/expand said CD34+CD38− hematopoictic progenitor cells.