Physiology of cultured animal cells
| Autor: | Lena Häggström, Jan Ljunggren, Magnus Doverskog, Lars Öhman |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
medicine.medical_treatment
Physiology Bioengineering Sf9 Spodoptera Biology Applied Microbiology and Biotechnology Mice Stress Physiological Tumor Cells Cultured medicine Animals Amino Acids Overflow metabolism Cells Cultured chemistry.chemical_classification Hybridomas Cell growth Growth factor General Medicine Metabolism Amino acid Glutamine Biochemistry chemistry Cell culture Energy Metabolism Multiple Myeloma Cell Division Biotechnology |
| Zdroj: | Journal of Biotechnology. 59:103-115 |
| ISSN: | 0168-1656 |
| DOI: | 10.1016/s0168-1656(97)00172-7 |
| Popis: | The physiology of cultured animal cells, in particular hybridoma, myeloma and insect cells, with respect to growth and proliferation, amino acid metabolism, energy metabolism and cellular responses to environmental stress is discussed in this paper. The rate of proliferation of hybridoma cells in serum-containing media is limited by growth factors at a surprisingly early stage of growth. To maintain exponential growth in a batch culture, it is necessary to stimulate cell proliferation with repeated additions of serum or pure growth factor. It is further suggested that proliferation of Spodoptera frugiperda (Sf9 insect cells), a normal cell line able to grow in a serum-free medium without any added growth factors, is regulated by autocrine growth factors and possibly by other regulatory mechanisms, as Sf9 cells secrete a growth factor (IGF-I) and the medium still appears nutritionally sufficient at the time of cessation of growth. The uptake and metabolism of amino acids is one of the determinants of growth and production. Wasteful overproduction of amino acids in myeloma and hybridoma cells is a result of excess glutamine, and can be avoided by glutamine limitation. Synthesis of amino acids may be conditional, as in Sf9 cells which synthesise glutamine provided that ammonium is supplied to the medium; and cysteine (from methionine) provided that a sufficiently young inoculum is used. Uptake of amino acids in Sf9 cells appears regulated in relation to the proliferative status as there is a distinct cessation of uptake even before growth ceases. The energy metabolism in myeloma, hybridoma and insect cells is a typically substrate-concentration-dependent overflow metabolism. Substrate limitation (glucose and glutamine) decreases by-product formation and increases metabolic efficiency in all these cell lines. However, glutamine limitation, as used in fed-batch cultures (or chemostat cultures) provokes cell death (in parallel to growth) in hybridoma cells in the concentration range below 0.05 mM. |
| Databáze: | OpenAIRE |
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