Optimization of human long-term bone marrow cultures.

TL;DRAbstract

Human long-term bone marrow cultures (LTBMCs) have been limited both in their longevity (8 to 12 weeks) and in their cell production over time. Hematopoiesis in vivo continues unabated throughout life, suggesting that failure to obtain continuous hematopoiesis in vitro is probably due to the failure of culture conditions to accurately reproduce the in vivo state. This thesis will investigate the hypothesis that the longevity and productivity of LTBMCs can be substantially improved by modelling the in vitro culture conditions on the in vivo bone marrow microenvironment. First, culture productivity and longevity in LTBMCs was found to be relatively independent of the variation in inoculum density from 1-5 $\cdot$ 10$\sp6$ cells/ml, but was significantly altered by the medium exchange rate. Increased culture exchange was superior to traditional Dexter schedules, with 1/2 medium volume exchange per day being the optimal. Second, neither collagen type 1 nor fibronectin coated tissue culture

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