Culturing cells on macroporous glass carriers coated with gelatin, extracellular matrix protein and stromal cells

摘要

Stem cells or other organ-function cells are cultivated in a fluidized bed system on macroporous glass carrier bodies treated with a structure protein such as gelatin and an extracellular matrix protein such as fibronectin, and coated with a stroma cell layer. Glass carriers coated with gelatin are added to a fluidized bed reactor, and a culture medium containing an extracellular matrix protein is added to bind the protein to the gelatin. Stromal cells are then added and the cells are cultured to immobilize the cells on the carriers containing the bound protein. Immature organ-function cells are added to the reactor, and while generating a fluidized bed of the carriers in the culture medium, the culture medium is recirculated from and to the reactor in a recirculation loop. Bubble-free aeration of the culture medium is effected to cultivate the immature organ-function cells on the carriers to obtain both mature differentiated organ-function cells and progenitor organ-function cells. Mature differentiated organ-function cells which are released from the carriers are harvested from the reactor while retaining the progenitor organ-function cells which adhere to the carriers to provide an equilibrium between the harvested mature differentiated organ-function cells, the progenitor cells and the immature cells added to the reactor. Growth and regulatory factors for cultivating organ-function cells can be produced by culturing immobilized stromal cells in the reactor in the absence of the organ-function cells.