Long-term cultivation of mammalian cell strains in protein- and lipid-free chemically defined synthetic media

Takaoka, T; Katsuta, H

doi:10.1016/0014-4827(71)90412-5

Cited by 68 publications

(12 citation statements)

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“…It became clear in later studies that trace elements contaminating the water or raw materials they used are necessary for the protein‐free cultivation of CHO cells in Ham's F‐12 medium, leading to the development of MCDB301, a medium that is supplemented with 20 trace elements (Table 2). 60 Toshiko Takaoka and Hajim Katsuta successfully cultured various kinds of cells long‐term in a simple medium that did not contain proteins or lipids;61 however, only a few cell types can adapt to protein‐ and lipid‐free conditions and the fatty‐acid composition of the cells thus cultured differs greatly from the cells cultured in serum‐containing media 62…”

Section: History Of Cell Culture Mediamentioning

confidence: 99%

Animal‐cell culture media: History, characteristics, and current issues

Yao

Asayama

2017

Reprod Medicine & Biology

374

278

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BackgroundCell culture technology has spread prolifically within a century, a variety of culture media has been designed. This review goes through the history, characteristics and current issues of animal‐cell culture media.MethodsA literature search was performed on PubMed and Google Scholar between 1880 and May 2016 using appropriate keywords.ResultsAt the dawn of cell culture technology, the major components of media were naturally derived products such as serum. The field then gradually shifted to the use of chemical‐based synthetic media because naturally derived ingredients have their disadvantages such as large batch‐to‐batch variation. Today, industrially important cells can be cultured in synthetic media. Nevertheless, the combinations and concentrations of the components in these media remain to be optimized. In addition, serum‐containing media are still in general use in the field of basic research. In the fields of assisted reproductive technologies and regenerative medicine, some of the medium components are naturally derived in nearly all instances.ConclusionsFurther improvements of culture media are desirable, which will certainly contribute to a reduction in the experimental variation, enhance productivity among biopharmaceuticals, improve treatment outcomes of assisted reproductive technologies, and facilitate implementation and popularization of regenerative medicine.

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Section: History Of Cell Culture Mediamentioning

confidence: 99%

Animal‐cell culture media: History, characteristics, and current issues

Yao

Asayama

2017

Reprod Medicine & Biology

374

278

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“…In order to circumvent these problems, several cell lines have been adapted to proliferate in serum-free media (1)(2)(3)(4) (5)(6)(7)(8).…”

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Growth of a rat neuroblastoma cell line in serum-free supplemented medium.

Bottenstein¹,

Sato²

1979

Proc. Natl. Acad. Sci. U.S.A.

1,945

983

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The rat neuroblastoma B104 cell line, which originated in the central nervous system, was able to proliferate in the absence of serum in synthetic medium supplemented with insulin, transferrin, progesterone, selenium, and putrescine. When added individually, each supplement had little or no effect; however, in combination there was a marked synergistic effect on cell number. The cells attained the same saturation density in this medium as in medium with 10% fetal calf serum added. More extensive process formation was observed in the supplemented medium, and other differentiated properties were retained as well. Synthetic media generally require serum supplementation to support the proliferation or survival of cultured cells. The inclusion of serum, however, may significantly affect experimental reproducibility, because batch variations occur as a result of differences in donor age, sex, nutrition, and physiological state even in pooled serum samples. In addition, the complex undefined nature of serum is a complication when assessing the effect(s) of regulatory agents, such as hormones or neurotransmitters, on differentiated properties of nervous system cells in culture. This is particularly important for long-term studies, because if serum is deleted a substantial reduction of cell numbers, and in many cases complete cell death, may occur within hours or a few days.In order to circumvent these problems, several cell lines have been adapted to proliferate in serum-free media (1-4). How The B104 rat neuroblastoma, a cell line of central nervous system origin, exhibits many of the properties characteristic of differentiated neurons, such as generation of action potentials, synthesis of neurotransmitters, and presence of neurotransmitter receptors and the neuron-specific 14-3-2 protein (9, 10). Furthermore, B104 cells, like C1300 neuroblastoma cells (11, 12), respond to removal of serum by rapidly extending neurites, a phenomenon that has been correlated with the preceding neuronal properties (9).In this communication we report that B104 cells can proliferate in a serum-free synthetic medium supplemented with insulin, transferrin, progesterone, selenium, and putrescine (N2 medium). More extensive process formation is seen in N2 than in serum-supplemented medium, and other differentiated properties have been retained as well. METHODS AND MATERIALSCell Culture. The rat neuroblastoma B104 cell line was obtained from D. Schubert of the Salk Institute, La Jolla, CA. Stock cultures were maintained in a 1:1 mixture of Ham's F12 medium and the Dulbecco-Vogt modification of Eagle's medium (DME) supplemented with 5% (vol/vol) horse serum; 2.5% (vol/vol) fetal calf serum; 1.2 g of NaHCO3 per liter; 15 mM Hepes buffer; and 40 mg of penicillin, 8 mg of ampicillin, and 90 mg of streptomycin per liter. Triple-distilled water was used to prepare media. Stock cultures were grown in flasks (Falcon Plastics 3013, 25-cm2 surface area) in 4 ml of medium in a humidified atmosphere of 5% C02/95% air at 370C and subcultured every ...

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“…The cells grew in proteinfree F-10 medium with a 30-hr population-doubling time. Intensive efforts have been made to develop human cell lines in a protein-free chemically defined medium (4,5). However, only a few human cells have been shown to proliferate in protein-free media for a long time.…”

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Long-term cultivation and differentiation of human erythroleukemia cells in a protein-free chemically defined medium.

Okabe¹,

Fujisawa²,

Takaku³

1984

Proc. Natl. Acad. Sci. U.S.A.

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To examine whether a human erythroleukemia cell line, K-562, can proliferate and differentiate without protein supplements, long-term cultivation of the cells was carried out in a protein-free chemically defined medium. By the use of stepwise decreases in the fetal bovine serum concentration, continuous growth of K-562 cells was established in a protein-free F-10 medium. The cells have been propagated in this medium for 3 years. The population-doubling time of the cells is about 30 hr. Growth was not stimulated by the addition of insulin, epidermal growth factor, fibroblast growth factor, multiplication-stimulating activity, transferrin, platelet-derived growth factor, or dexamethasone. Addition of serum stimulated the cell growth slightly and increased the saturation density. The saturation density of the cells could be increased to that seen with serum-supplemented cultures by changing the serum-free medium daily. The cells synthesized significant amounts of hemoglobin in the presence of hemin without serum supplementation. The results suggest that the human erythroleukemia cells grown in a protein-free medium do not require serum components for their growth or hemin-induced hemoglobin synthesis and provide an excellent model for better understanding of the growth and differentiation of human leukemia cells.Learning how cells of multicellular animals control their proliferation rates has long been a key objective of experimental biology. Nutritionally simple microbes offer several advantages for biosynthetic studies on animal tissues. Compared to Escherichia coli, our knowledge of the structural organization and biochemistry of eukaryotic cells is still pitifully meager. The greater potential of tumor cells to yield cell lines has led to the belief that all cells that grow as cell lines perhaps do so because they have acquired many of the essential properties of the cancer cell by mutation(s). In general, the growth in culture requires not only a well-defined collection of amino acids and vitamins but also depends upon protein factors present in blood serum. In the absence of such growth-promoting factors, most cells enter a resting state of G1 phase of cell cycle, with a substantial reduction of cell numbers or complete cell death within a few days. The mechanisms by which such factors affect cell functions have not been understood fully. Among these factors, even the effects of insulin on cell growth and functions have not been clarified (1). The complex nature of such factors represents an undefined variable when assessing the biochemical changes within the cell during proliferation or differentiation. To circumvent this problem, long-term cultivation of human erythroleukemia cells was performed in a protein-free chemically defined medium. This protein-free culture may provide advantages similar to bacterial cultures, especially for genetic research of cells, because mutations to auxotrophy in this system are possible in the absence of serum factors. MATERIALS AND METHODSCell Culture. A human erythr...

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Long-term cultivation of mammalian cell strains in protein- and lipid-free chemically defined synthetic media

Cited by 68 publications

References 13 publications

Animal‐cell culture media: History, characteristics, and current issues

Animal‐cell culture media: History, characteristics, and current issues

Growth of a rat neuroblastoma cell line in serum-free supplemented medium.

Long-term cultivation and differentiation of human erythroleukemia cells in a protein-free chemically defined medium.

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