Physiology of cultured animal cells
Physiology of cultured animal cells
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DOI：
10.1016/s0168-1656(97)00172-7
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发表时间：
1997-12-17
影响因子：
4.1
通讯作者：
Häggström, L
中科院分区：
文献类型：
--
作者：
Doverskog, M;Ljunggren, J;Häggström, L
本文讨论了培养的动物细胞，特别是杂交瘤细胞、骨髓瘤细胞和昆虫细胞在生长和增殖、氨基酸代谢、能量代谢以及细胞对环境胁迫的反应等方面的生理学。杂交瘤细胞在含血清培养基中的增殖速率在令人惊讶的早期生长阶段受到生长因子的限制。为了在分批培养中保持指数生长，有必要通过重复添加血清或纯生长因子来刺激细胞增殖。进一步表明，草地贪夜蛾（Spodoptera frugiperda）（Sf 9昆虫细胞）（一种能够在没有任何添加的生长因子的无血清培养基中生长的正常细胞系）的增殖受到自分泌生长因子的调节，并且可能受到其他调节机制的调节，因为Sf 9细胞分泌生长因子（IGF-I），并且培养基在生长停止时仍然表现出营养充足。氨基酸的吸收和代谢是生长和生产的决定因素之一。在骨髓瘤和杂交瘤细胞中浪费性的过量生产氨基酸是过量谷氨酰胺的结果，并且可以通过限制谷氨酰胺来避免。氨基酸的合成可以是有条件的，如在Sf 9细胞中，其合成谷氨酰胺，条件是向培养基中供应铵;和半胱氨酸（来自甲硫氨酸），条件是使用足够年轻的接种物。Sf 9细胞中氨基酸的摄取似乎与增殖状态有关，因为即使在生长停止之前也有明显的摄取停止。骨髓瘤、杂交瘤和昆虫细胞的能量代谢是典型的底物浓度依赖性溢流代谢。底物限制（葡萄糖和谷氨酰胺）减少了副产物的形成，并增加了所有这些细胞系的代谢效率。然而，谷氨酰胺限制，如在补料分批培养（或恒化培养）中使用的，在低于0.05 mM的浓度范围内引起杂交瘤细胞的细胞死亡（与生长平行）。
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. (C) 1997 Elsevier Science B.V.