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血清补充剂和无血清培养基:在动物细胞微载体培养中的适用性。

Serum supplements and serum-free media: applicability for microcarrier culture of animal cells.

作者信息

Clark J M, Gebb C, Hirtenstein M D

出版信息

Dev Biol Stand. 1981;50:81-91.

PMID:7341300
Abstract

Several different combinations of serum supplements and also serum-free media were examined for their ability to support the growth of MRC-5 and Vero cells on Cytodex microcarriers. Greater economy of serum was achieved for routine microcarrier culture by selecting the type of serum supplement, on the basis of whether the supplement was to support attachment and growth of cells at low densities or growth of cells at later stages in the culture cycle. Further economy was achieved by altering the serum concentration according to the requirements of each stage of the culture cycle. More consistent results were obtained when batches of serum were selected on the basis of quality control tests with microcarrier culture as well as with monolayer culture. Low serum and serum-free media were able to support the growth of cells in microcarrier cultures. A mixture of DME/Medium 199 (50/50) containing foetal calf serum (0.5%, v/v), BSA and EGF supported growth of MRC-5 and Vero cells to nearly the same extent as DME/Medium 199 containing 10% (v/v) foetal calf serum. Serum-free media was used to achieve cell yields at least one half of those attained with serum-supplemented media. Maximum growth was obtained when DME/Medium 199 was supplemented with fibronectin, BSA, insulin, transferrin, putrescine, fetuin and EFG. Fetuin was omitted and replaced by dexamethasone and trace metals with only a slight reduction in cell yield. Fibronectin was essential in all serum-free media.

摘要

研究了几种不同的血清补充剂组合以及无血清培养基,以考察它们在Cytodex微载体上支持MRC - 5和Vero细胞生长的能力。通过根据血清补充剂是支持低密度细胞贴壁和生长还是支持培养周期后期细胞生长来选择血清补充剂类型,可在常规微载体培养中实现更高的血清经济性。根据培养周期各阶段的需求改变血清浓度可进一步实现经济性。当根据微载体培养以及单层培养的质量控制测试来选择血清批次时,可获得更一致的结果。低血清和无血清培养基能够支持微载体培养中的细胞生长。含有胎牛血清(0.5%,v/v)、牛血清白蛋白和表皮生长因子的DME/199培养基(50/50)混合物支持MRC - 5和Vero细胞生长的程度与含有10%(v/v)胎牛血清的DME/199培养基几乎相同。使用无血清培养基获得的细胞产量至少是含血清培养基的一半。当DME/199培养基补充纤连蛋白、牛血清白蛋白、胰岛素、转铁蛋白、腐胺、胎球蛋白和表皮生长因子时可获得最大生长。省略胎球蛋白并用地塞米松和痕量金属替代,细胞产量仅略有降低。在所有无血清培养基中纤连蛋白都是必需的。

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