Methods, compositions and devices for growing human hematopoietic cells

A method of growing human stem cells in culture, said method comprising:

• inoculating a reactor vessel comprising, on a surface, heterologous stromal cells adherent to one side of a protein membrane with pores in a range of about 1-5 microns with human hematopoietic cells comprising human stem cells found in the human hematopoietic system, said inoculation being on the opposite side of said membrane from said stromal cells, wherein at least a portion of said stromal cells are transformed and capable of excreting a colony stimulating factor or an interleukin which directs the proliferation, differentiation, or both, of said human stem cells;
• substantially continuously perfusing said cells in said reactor with a liquid culture medium comprising any additional growth factors necessary for proliferation, differentiation, or both, of said human hematopoietic cells, while maintaining said reactor under physiologically acceptable conditions, said cells being perfused at a rate providing ex vivo proliferation, differentiation, or both, of said human stem cells;
• and
• harvesting human hematopoietic cells from said reactor.

A method of growing human stem cells in culture, said method comprising:

• inoculating a reactor vessel comprising, on a surface, stromal cells adherent to a protein substrate, with human hematopoietic cells comprising human stem cells found in the human hematopoietic system, wherein at least a portion of said stromal cells are transformed and capable of excreting at least one growth factor which directs the proliferation, differentiation, or both, of said human stem cells;
• substantially continuously perfusing said cells in said reactor with a liquid culture medium comprising any additional growth factors necessary for proliferation, differentiation, or both, of said human stem cells, while maintaining said reactor under physiologically acceptable conditions, said cells being perfused either at a rate providing ex vivo division of said human stem cells or at a rate sufficient to maintain production of hematopoietic growth factors at about the endogenous level produced by said normal human bone marrow stromal cells;
• and
• harvesting human hematopoietic cells from said reactor,
• with the proviso that when said human hematopoietic cells inoculated into said reactor vessel are suspected of comprising neoplastic cells, said perfusing is at rate providing a force on said cells greater than the affinity of neoplastic cells to said stromal cells and less than the affinity of normal human hematopoietic cells to said stromal cells.

A method of growing human hematopoietic cells in culture, said method comprising:

• inoculating a reactor vessel comprising, on a surface, heterologous stromal cells adherent to one side of a protein substrate with pores in the range of about 1-5 microns with human hematopoietic cells comprising human progenitor cells found in the human hematopoietic system, said inoculation being on the opposite side of said membrane from said stromal cells, wherein at least a portion of said stromal cells are transformed fibroblast cells capable of adhering to a protein substrate and capable of excreting at least one growth factor which directs proliferation, differentiation, or both, of said human progenitor cells;
• continuously or periodically perfusing said cells in said reactor with a liquid culture medium comprising any additional growth factors necessary for proliferation, differentiation, or both, of said human hematopoietic progenitor cells, while maintaining said reactor under physiologically acceptable conditions, said cells being perfused at a rate equal to a rate of from 50 to 100% daily liquid culture medium replacement for a cell density of from 1.times.10.sup.4 to 1.times.10.sup.7 cells per ml of culture;
• and
• harvesting hematopoietic cells from said reactor, with the proviso that when said human hematopoietic cells inoculated into said reactor vessel are suspected of comprising neoplastic cells, said perfusing is at a rate providing a force on said cells greater than the affinity of neoplastic cells to said stromal cells and less than the affinity of normal hematopoietic cells to said stromal cells.