An improved heat‐stable glutamine‐free chemically defined medium for growth of mammalian cells

Nagle, Stanley C.; Brown, Bruce L.

doi:10.1002/jcp.1040770214

Cited by 32 publications

(5 citation statements)

References 12 publications

(1 reference statement)

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“…For example, Eagle (1959) noted that cell cultures of carcinosarcoma, rabbit fibroblasts and embryonic limb-bud cells, and monkey kidney cells have unusual requirements for ASN, SER, and GLY, respectively (Eagle et al, 1958). Similarly, ascites tumor cells utilize LEU at very high rates (Lazo, 1981); and with L-cells, ALA is essential in the absence of GLN (Nagle and Brown, 1971). In the present studies, growth plate chondrocytes catabolized GLN very rapidly (Figs.…”

Section: Discussionmentioning

confidence: 70%

Utilization and formation of amino acids by chicken epiphyseal chondrocytes: Comparative studies with cultured cells and native cartilage tissue

Ishikawa

Chin

Hubbard

et al. 1985

Journal Cellular Physiology

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Utilization and production of amino acids by primary cultures of chicken growth plate epiphyseal chondrocytes grown in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum were investigated in both short-term (6-72 h) and long-term (3-24 day) cultures. Comparative studies were made on levels of free amino acids in chicken blood plasma and serum, and in extracellular fluids from different regions of growth plate cartilage and from two types of muscle. Chondrocytes rapidly consumed glutamine from the medium, and to lesser extents, various other amino acids. In contrast, free ammonia, alanine, glycine, glutamate, proline, and aspartate were released into the medium. The utilization of certain amino acids changed, depending on the stage of culture. Initially glutamate was released into the medium but after confluency was consumed. Conversely, histidine, lysine, and phenylalanine were initially utilized but later were released into the medium. Levels of total free amino acids in extracellular fluids of cartilage and muscle were higher than those in plasma and serum, while in cartilage the levels increased progressively from the resting to the hypertrophic zones. In these sequential regions certain amino acids increased proportionally, whereas others decreased. These interrelationships generally correlated closely with metabolism of amino acids by the cultured chondrocytes. They indicate that significant differences in amino acid metabolism exist between tissue areas and are reflected in the extracellular fluid composition. Accordingly, adjustment of specific amino acids may optimize culture conditions, enabling more normal phenotypic expression in vitro.

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Section: Discussionmentioning

confidence: 70%

Utilization and formation of amino acids by chicken epiphyseal chondrocytes: Comparative studies with cultured cells and native cartilage tissue

Ishikawa

Chin

Hubbard

et al. 1985

Journal Cellular Physiology

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“…With two exceptions, all of the additives in L-15 growth medium (except Methocel, which was never included) were present. For experiments with [ 3H]glutamate, glutamine was omitted from the medium ; in two cases, alanine (which can substitute for glutamine as an amino donor ; Nagle and Brown, 1971) was used in place of glutamine . Preliminary experiments with [ 14 C]histidine indicated that imidazole might compete with histidine for uptake, so glycerol-2-phosphate was used as the buffer in experiments with [ 14C]histidine.…”

Section: Incubations With Radioactivementioning

confidence: 99%

Primary Cultures of Dissociated Sympathetic Neurons

Mains

Patterson

1973

The Journal of Cell Biology

400

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Rat sympathetic ganglia were disrupted by mechanical agitation to yield dissociated primary neurons, and the conditions for long-term growth in culture of the isolated neurons were examined. The neurons were grown with or without non-neural cells, simply by the addition or deletion of bicarbonate during growth in culture. Fluorescence histochemistry indicated that the isolated neurons contained catecholamines; incubations with radioactive precursors were used to verify the synthesis and accumulation of both dopamine and norepinephrine. The neurons also produced octopamine using tyramine as precursor, but not with tyrosine as the precursor. In the presence of eserine, older cultures synthesized and stored small amounts of acetylcholine. The cultures did not synthesize and accumulate detectable levels of radioactive γ-aminobutyric acid, 5-hydroxytryptamine, or histamine.

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“…Because heat sterilization is likely a more efficient process at industrial scale, formulation of cultivated meat media using more heat stable ingredients should be considered, though this idea has not yet been directly tested. Heat stable, defined serum‐free media have been developed for other cell types and applications (Nagle & Brown, 1971), which lends credibility to the notion that this may be possible for cultivated meat.…”

Section: Common Media Formulations For Animal Cell Culture and Issuesmentioning

confidence: 97%

Considerations for the development of cost‐effective cell culture media for cultivated meat production

O’Neill

Cosenza

Baar

et al. 2020

Comp Rev Food Sci Food Safe

116

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Innovation in cultivated meat development has been rapidly accelerating in recent years because it holds the potential to help attenuate issues facing production of dietary protein for a growing world population. There are technical obstacles still hindering large‐scale commercialization of cultivated meat, of which many are related to the media that are used to culture the muscle, fat, and connective tissue cells. While animal cell culture media has been used and refined for roughly a century, it has not been specifically designed with the requirements of cultivated meat in mind. Perhaps the most common industrial use of animal cell culture is currently the production of therapeutic monoclonal antibodies, which sell for orders of magnitude more than meat. Successful production of cultivated meat requires media that is food grade with minimal cost, can regulate large‐scale cell proliferation and differentiation, has acceptable sensory qualities, and is animal ingredient‐free. Much insight into strategies for achieving media formulations with these qualities can be obtained from knowledge of conventional culture media applications and from the metabolic pathways involved in myogenesis and protein synthesis. In addition, application of principles used to optimize media for large‐scale microbial fermentation processes producing lower value commodity chemicals and food ingredients can also be instructive. As such, the present review shall provide an overview of the current understanding of cell culture media as it relates to cultivated meat.

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An improved heat‐stable glutamine‐free chemically defined medium for growth of mammalian cells

Cited by 32 publications

References 12 publications

Utilization and formation of amino acids by chicken epiphyseal chondrocytes: Comparative studies with cultured cells and native cartilage tissue

Utilization and formation of amino acids by chicken epiphyseal chondrocytes: Comparative studies with cultured cells and native cartilage tissue

Primary Cultures of Dissociated Sympathetic Neurons

Considerations for the development of cost‐effective cell culture media for cultivated meat production

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