LARGE-SCALE PRODUCTION OF MURINE BONE-MARROW CELLS IN AN AIRLIFT PACKED-BED BIOREACTOR

Large-scale cultivation of murine bone marrow cells was accomplished i n an airlift packed bed bioreactor system designed to mimic the in viv o bone marrow environment. The attachment-dependent stromal cell popul ation, which provides the necessary microenvironment, including growth factors for subsequent hematopoietic activity, was first established within the bioreactor. This attachment-dependent cell growth occurred on the fiberglass matrix packed in the annular region of the bioreacto r. Once the stromal cell layer was established, fresh bone marrow cell s were inoculated to initiate hematopoiesis. However, traditional cult ure medium was found to be inadequate for the initiation of hematopoie sis, but the use of stromal cell ''conditioned'' medium (with no exoge nously added growth factors) yielded sustained cell production. The ex tent of stromal cell subculturing prior to inoculation into the biorea ctor and the inoculation density were also important factors for the s uccessful initiation of hematopoietic activity. A 500-mL perfusion cul ture experiment resulted in the production and harvest of 3.6 x 10(8) suspended bone marrow cells over the course of 11 weeks. (C) 1996 John Wiley & Sons, Inc.