Hematopoietic cell culture therapies (Part I): cell culture considerations

PER.C6(R) cell growth, metabolism, and adenovirus production were studied in head-to-head comparisons in stirred bioreactors under different pH conditions. Cell growth rate was found to be similar in the pH range of 7.1-7.6, while a long lag phase and a slower growth rate were observed at pH 6.8. The specific consumption rates of glucose and glutamine decreased rapidly over time during batch cell growth, as did the specific lactate and ammonium production rates. Cell metabolism in both infected and uninfected cultures was very sensitive to culture pH, resulting in dramatic differences in glucose/glutamine consumption and lactate/ammonium production under different pH conditions. It appeared that glucose metabolism was suppressed at low pH but the efficiency of energy production from glucose was enhanced. Adenovirus infection resulted in profound changes in cell growth and metabolism. Cell growth was largely arrested under all pH conditions, while glucose consumption and lactate production were elevated post virus infection. Virus infection induced a reduction in glutamine consumption at low pH but an increase at high pH. The optimal pH for adenovirus production was found to be 7.3 under the experimental conditions used in the study. Deviations from this optimum resulted in significant reductions of virus productivity. The results indicate that culture pH is a very critical process parameter in PER.C6(R) cell culture and adenovirus production.

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“…65 and 6.8 (LeBoeuf and Kerckaert, 1986). Human hematopoietic differentiation was also found to be sensitive to pH (McAdams et al, 1996).…”

Section: Introductionmentioning

confidence: 96%

Serum‐free suspension cultivation of PER.C6® cells and recombinant adenovirus production under different pH conditions

Xie

Pilbrough

Metallo

et al. 2002

Biotech & Bioengineering

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“…Generally, HSPCs expanded ex vivo cannot be used in clinics due to their biological incompatibility. It is known that oxygen concentration is one of the important factors for proliferation and differentiation of HSPCs ex vivo (McAdams et al, 1996). For cultivation ex vivo, low levels of oxygen favored the proliferation of colony-forming cells (CFCs) (Broxmeyer et al, 1990;Rich and Kubanek, 1982), HSPCs (Koller et al, 1992a,b), and mature granulocytes (Hevehan et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Role of the plasma membrane ROS-generating NADPH oxidase in CD34+ progenitor cells preservation by hypoxia

Fan

Cai

Tan

2007

Journal of Biotechnology

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Hypoxia favored the preservation of progenitor characteristics of hematopoietic stem and progenitor cells (HSPCs) in bone marrow. This work aimed at studying the role of reactive oxygen species (ROS)-generating NADPH oxidase system regulated by hypoxia in ex vivo cultures of cord blood CD34 + cells. The results showed that NADPH oxidase activity and ROS generation were reduced in hypoxia with respect to normal oxygen tension. Meanwhile the ROS generation was found to be inhibited by diphenyleneiodonium (the NADPH oxidase inhibitor), or N-acetylcysteine (the ROS scavenger). Accordingly NADPH oxidase mRNA and p67 protein levels decreased in hypoxia. The analysis of progenitor characteristics, including the proportion of cultured cells expressing the HSPCs marker CD34 + CD38 − , colony production ability of the colony-forming cells (CFCs), and the re-expansion capability of the cultured CD34 + cells, showed that either 5% pO 2 or reduced ROS favored preserving the characteristics of CD34 + progenitors, and promoted the expansion of CD34 + CD38 − cells as well. The above results demonstrated that hypoxia effectively maintained biological characteristics of CD34 + cells through keeping lower intracellular ROS levels by regulating NADPH oxidase.

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“…The lifespan of cells in culture is likely to depend on cytokine stimulation as well as on a number of physicochemical parameters, such as pH, dissolved oxygen, and nutrient and metabolite concentrations (McAdams et al, 1996a).…”

Section: Introductionmentioning

confidence: 99%

Real-time method for determining the colony-forming cell content of human hematopoietic cell cultures

Collins

Nielsen²,

Wong³

et al. 1997

Biotechnol. Bioeng.

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Hematopoietic cell culture therapies (Part I): cell culture considerations

Cited by 48 publications

References 38 publications

Serum‐free suspension cultivation of PER.C6® cells and recombinant adenovirus production under different pH conditions

Serum‐free suspension cultivation of PER.C6® cells and recombinant adenovirus production under different pH conditions

Role of the plasma membrane ROS-generating NADPH oxidase in CD34+ progenitor cells preservation by hypoxia

Real-time method for determining the colony-forming cell content of human hematopoietic cell cultures

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