To provide a novel method, including culture media conditions, reactors, etc., for ex vivo human stem cell proliferation and transformation of stable genes, as well as a novel method, including culture media conditions and reactors, for optimum conditions for (i) culture of human hematopoietic precursor cells and (ii) transformation of stable genes of human hematopoietic precursor cells.
Disclosed is a culture method for obtaining ex vivo human stem cell division products derived from hepatopoietic system featured by that cell compositions including human hepatopoietic system-derived stem cells are cultured in a liquid culture medium which is continuously or periodically replaced at a rate of 1 mL of medium per 1 mL of culture for about 24 to about 48 hours, metabolic products are removed while maintaining the culture under physiologically acceptable conditions and depleted nutrients are replenished. According to the present invention, human stem cells are efficiently obtained ex vivo by culturing human stem cells and/or human hematopoietic precursor cells and/or human stromal cells in a liquid culture medium which is continuously or periodically replaced, preferably perfused, and replenishing nutrients while maintaining the culture under physiologically acceptable conditions.
CLARKE MICHAEL F
PALSSON BERNHARD O
SCHWARTZ RICHARD M
Toshio Takano
Toshio Nakajima
Hiromi Matoba