Bioreactor for Mammalian Cell Culture

Prokop, A.; Rosenberg, Morris Z.

doi:10.1515/9783112536827-002

Cited by 2 publications

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“…1f). The importance of carefully controlling the environmental conditions is, however, not a new realization: recommendations made over three decades ago included the use of chemostats 3 and bioreactors 4 for improving the monitoring and control of culture conditions.…”

Section: Best Practicesmentioning

confidence: 99%

“…However, the culture methods have remained relatively unchanged. Major advances in culture systems were made over three decades ago 3,4 , yet the old standard approach of batch cell culturethe culture of cells either in suspension or as adherent monolayers of cells in standard media [5][6][7] remains the predominant method in biomedical research.Culture media provides crucial nutrients, signalling molecules (such as growth factors), and suitable osmotic conditions. The gaseous and thermal environments of cell cultures are typically controlled by the incubator.…”

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A prevalent neglect of environmental control in mammalian cell culture calls for best practices

et al. 2021

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Human cell lines, first cultured in the 1950s 1 , are indispensable in biomedical research. Today, a wide range of cell types are available, and sophisticated advanced 'omics' and visualization techniques allow for the routine assessment of cell identity and cellular responses 2 . However, the culture methods have remained relatively unchanged. Major advances in culture systems were made over three decades ago 3,4 , yet the old standard approach of batch cell culturethe culture of cells either in suspension or as adherent monolayers of cells in standard media [5][6][7] remains the predominant method in biomedical research.Culture media provides crucial nutrients, signalling molecules (such as growth factors), and suitable osmotic conditions. The gaseous and thermal environments of cell cultures are typically controlled by the incubator. The initial media conditions are generally stabilized by adjusting them to 18.6% O2 and a standard pH of 7.4, and this adjustment is achieved by adding a given amount of HCO3 − salt (a base), and by enriching the media with CO2 to a given percentage in the air (usually 5% or 10%). However, cell metabolism involves the exchange of gasesspecifically the release of CO2 and the consumption of O2and this can affect cellular growth via the alteration of, for example, the pH and the level of dissolved O2 (dO2) in the cellular microenvironment 8 . In theory, the equilibration of the medium with the gaseous and thermal environments of the incubator provides a way to reliably mimic O2, CO2 and HCO3 − homeostasis in metazoan body fluids. Yet this doesn't take into account the fact that homeostasis in a living mammal is supported by the active exchange of gases with the atmosphere. The absence of such active gas exchange in cell cultures suggests that, over time, cellular metabolic activities might acidify and deoxygenate the cellular microenvironment 8-10 , if intermittent monitoring and (when necessary) corrective action are not carried out.To mimic a physiological environment when using cell cultures, careful control over environmental factors (such as pH, CO2 and O2) is typically needed, in particular because even small deviations of environmental parameters from physiological levels may impair cellular function. For instance, in human blood, pH values below 7.2 (acidaemic conditions) and above 7.44 (alkalaemic conditions) can be fatal [11][12][13] . In cell cultures, the optimal growth of normal cells (that is, non-cancerous cells and non-transformed cells) occurs within a specific alkaline pH range, whereas cancer cells grow in a broader pH range that is shifted towards acidic values [14][15][16][17][18] . Cells have evolved mechanisms, including the use of Na + /H + antiporters or histone deacetylation, that restore the alkaline pH of the cytoplasm when the extracellular pH deviates from physiological levels [19][20][21][22][23][24][25] . However, such regulatory mechanisms require cellular energy, and changes in the acetylation state of chromatin can alter gene transcription and reduc...

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Section: Best Practicesmentioning

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A prevalent neglect of environmental control in mammalian cell culture calls for best practices

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“…pH, temperature, oxygen concentrations), monitoring and supplying nutrients, removing waste products, and ensuring homogenous distribution through intermittent or continuous stirring, mammalian cell cultures up to 2 x 10 4 L have been reported, although no protocols have been published in the cultured meat field that achieve similar dimensions. [74][75][76][77][78] In addition, most adult stem cells including SCs and FAPs are adherent cell types. Therefore, protocols to grow these cells in vessels such as bioreactors or spinner flasks usually require the use of microcarriers.…”

Section: Proliferationmentioning

confidence: 99%

Decoding cultured meat production

Messmer¹

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Bioreactor for Mammalian Cell Culture

Cited by 2 publications

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A prevalent neglect of environmental control in mammalian cell culture calls for best practices

A prevalent neglect of environmental control in mammalian cell culture calls for best practices

Decoding cultured meat production

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