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Ann Biomed Eng. 1996 May-Jun;24(3):373-81.

Determination of specific oxygen uptake rates in human hematopoietic cultures and implications for bioreactor design.

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1
Department of Chemical Engineering, University of Michigan, Ann Arbor, USA.

Abstract

Oxygen plays an important role in the cultivation of primary cells ex vivo. In this study, we used hermetically sealed tissue culture well inserts equipped with oxygen electrodes to measure the oxygen utilization of cultured human bone marrow mononuclear cells (BM MNCs). The oxygen uptake rate (OUR) of BM MNCs was determined during a 14-day culture in which both adherent and nonadherent cells were present. Early in the culture, the cells exhibited very low OURs. The specific OURs (uptake rate per cell) were at approximately 0.005 mumol/10(6) cells/hr shortly after the initiation of culture. The OUR then increased as the cultures developed. After about 8 to 10 days of cultivation the specific OURs had increased to 0.038 +/- 0.006 and 0.025 +/- 0.005 mumol/10(6) cells/hr for adherent and nonadherent cells, respectively, after which no further increase was observed. Based on these oxygen uptake rate data, a mathematical model of oxygen diffusion was formulated and use to investigate issues associated with hematopoietic bioreactor design, including initial cell density, medium depth, reactor configuration, and oxygen partial pressure. In situ OUR measurements confirmed predicted oxygen limitations based on the mathematical model and the experimentally determined OURs. High-density hematopoietic cultures present design challenges in terms of sufficient and uniform delivery of oxygen to an active hematopoietic culture. These challenges can be met by using parallel-plate bioreactors with thin liquid layers.

PMID:
8734058
[Indexed for MEDLINE]
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