Journal: Journal of biological engineering

Article Title: Scale-up of CHO cell cultures: from 96-well-microtiter plates to stirred tank reactors across three orders of magnitude.

doi: 10.1186/s13036-024-00475-8

Figure Lengend Snippet: Fig. 1 Oxygen transfer rate (OTR) of CHO DP12 cell cultures. Two independent experiments (1 and 2) were performed. For both experiments, a round 96-deep-well microtiter plate and 250 mL shake flasks were inoculated. The µTOM device was used for online monitoring of the microtiter plates (dark red line and circles; light red line and triangles) and the TOM device for the shake flasks (black line and squares; grey line and diamonds). For clarity, only every 24th measuring point over time is marked as a symbol. The microtiter plate experiments were performed in 72 (Experiment 1) and 66 (Experiment 2) replicates and the shake flask experiments in 3 replicates each. For clarity, the low standard deviations are not shown in this figure but can be found in Fig. S2. Culture conditions TOM device: 250 mL glass flasks, temperature (T) = 36.5 °C, shaking frequency (n) = 140 rpm, shaking diameter (d0) = 50 mm, filling volume (VL) = 50 mL, 5% CO2, 70% rel. hum., medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1. Culture conditions µTOM device: round 96-deep-well microtiter plate, temperature (T) = 36.5 °C, shaking frequency (n) = 850 rpm, shaking diameter (d0) = 3 mm, filling volume (VL) = 1 mL, 5% CO2, humidified, medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1

Article Snippet: Cells of the suspension-adapted CHO DP12 cell line (clone#1934, ATCC CRL-12445) were stored in the vapor phase of liquid nitrogen.

Techniques:

Journal: Journal of biological engineering

Article Title: Scale-up of CHO cell cultures: from 96-well-microtiter plates to stirred tank reactors across three orders of magnitude.

doi: 10.1186/s13036-024-00475-8

Figure Lengend Snippet: Fig. 2 Oxygen transfer rate (OTR) of CHO DP12 cell cultures monitored by the µTOM device. A OTR curves of cultivations in a round 96-deep-well plate with different filling volumes (N = 8 for each filling volume). B OTR curves of cultivations in a square 96-deep-well plate with different filling volumes (N = 3 or 6 for each filling volume). For clarity, only every 24th measuring point over time is marked as a symbol. The standard deviations are shown as shaded areas. Culture conditions: temperature (T) = 36.5 °C, shaking frequency (n) = 850 rpm, shaking diameter (d0) = 3 mm, varying filling volume (VL), 5% CO2, humidified, medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1

Article Snippet: Cells of the suspension-adapted CHO DP12 cell line (clone#1934, ATCC CRL-12445) were stored in the vapor phase of liquid nitrogen.

Techniques:

Journal: Journal of biological engineering

Article Title: Scale-up of CHO cell cultures: from 96-well-microtiter plates to stirred tank reactors across three orders of magnitude.

doi: 10.1186/s13036-024-00475-8

Figure Lengend Snippet: Fig. 3 CHO DP12 cell cultivations in round 96-deep-well plates (dark and light red lines/circles and upward triangle), shake flasks (black and gray lines/squares and diamonds), and a stirred tank reactor (STR, blue line and pentagon). A Depicted is the oxygen transfer rate (OTR). The curves of the microtiter plate and shake flask cultivations are already shown in Fig. 1 and plotted here again for improved comparability. The data for the STR are interpolated over 3 h. The calculated OTR values between days 3 and 4 are distorted by a short-term failure of aeration and stirring and were therefore excluded from the data. For original data refer to Fig. S5 A. The dissolved oxygen tension (DOT) (green line and sideward triangle) of the stirred tank reactor is also plotted. For clarity, only one measuring point per day is plotted. B Displayed are the viable cell densities (VCD) and viabilities for all cultivations. Culture conditions TOM device: 250 mL glass flasks, temperature (T) = 36.5 °C, shaking frequency (n) = 140 rpm, shaking diameter (d0) = 50 mm, filling volume (VL) = 50 mL, 5% CO2, 70% rel. hum., medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1. Culture conditions µTOM device: round 96-deep-well microtiter plate, temperature (T) = 36.5 °C, shaking frequency (n) = 850 rpm, shaking diameter (d0) = 3 mm, filling volume (VL) = 1 mL, 5 % CO2, humidified, medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1. Culture conditions stirred tank reactor: 1.5 L reactor, temperature (T) = 36.5 °C, stirrer speed (n) = 360 rpm (Rushton turbine), filling volume (VL) = 600 mL, 5% CO2, aeration = 0.2 vvm (sparged), medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1

Article Snippet: Cells of the suspension-adapted CHO DP12 cell line (clone#1934, ATCC CRL-12445) were stored in the vapor phase of liquid nitrogen.

Techniques:

Journal: Journal of biological engineering

Article Title: Scale-up of CHO cell cultures: from 96-well-microtiter plates to stirred tank reactors across three orders of magnitude.

doi: 10.1186/s13036-024-00475-8

Figure Lengend Snippet: Fig. 4 CHO DP12 cell cultivations in round 96-deep-well plates (dark and light red lines/circles and upward triangle), shake flasks (black and gray lines/squares and diamonds), and a stirred tank reactor (blue line and pentagon). A Depicted is the oxygen transfer rate (OTR). The curves of the microtiter plate and shake flask cultivations are already shown in Fig. 1 and plotted here again for improved comparability. Additionally, the curve of a shake flask cultivation with increased volumetric power input (P/V) from Neuss et al. [34] is shown. The data for the STR are interpolated over 3 h. For original data refer to Fig. S5 B. For clarity, only one measuring point per day is shown. B Displayed are the viable cell density (VCD) and viability for all cultivations. Culture conditions TOM device: 250 mL glass flasks, temperature (T) = 36.5 °C, shaking frequency (n) = 140 rpm, shaking diameter (d0) = 50 mm, filling volume (VL) = 50 mL, 5% CO2, 70% rel. hum., medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1. Culture conditions µTOM device: round 96-deep-well microtiter plate, temperature (T) = 36.5 °C, shaking frequency (n) = 850 rpm, shaking diameter (d0) = 3 mm, filling volume (VL) = 1 mL, 5% CO2, humidified, medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1. Culture conditions stirred tank reactor: 1.5 L reactor, temperature (T) = 36.5 °C, stirrer speed (n) = 100–250 rpm (Rushton turbine), filling volume (VL) = 600 mL, 5% CO2, aeration = 0.2 vvm (sparged), medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1

Article Snippet: Cells of the suspension-adapted CHO DP12 cell line (clone#1934, ATCC CRL-12445) were stored in the vapor phase of liquid nitrogen.

Techniques:

Journal: Journal of biological engineering

Article Title: Scale-up of CHO cell cultures: from 96-well-microtiter plates to stirred tank reactors across three orders of magnitude.

doi: 10.1186/s13036-024-00475-8

Figure Lengend Snippet: Fig. 5 Offline measured metabolite and product concentrations of the cultivations shown in Fig. 4. CHO DP12 cell cultivations were performed in round 96-deep-well plates (dark and light red lines/circles and upward triangle), shake flasks (black and gray lines/squares and diamonds), and a stirred tank reactor (blue line and pentagon). Depicted are A) glutamine concentrations B) glucose concentrations C) lactate concentrations and D) IgG antibody titer

Article Snippet: Cells of the suspension-adapted CHO DP12 cell line (clone#1934, ATCC CRL-12445) were stored in the vapor phase of liquid nitrogen.

Techniques:

Journal: New biotechnology

Article Title: Impact of hydromechanical stress on CHO cells' metabolism and productivity: Insights from shake flask cultivations with online monitoring of the respiration activity.

doi: 10.1016/j.nbt.2024.09.008

Figure Lengend Snippet: Fig. 1. Cultivation of CHO DP12 cells in shake flasks under standard condi tions. The experiments were performed in six replicates. Three replicates were sampled daily for offline analysis and three were used for online monitoring only. All data are shown as mean value of three biological replicates (indicated as error bars or shades). A) Depicted are the viable cell density (VCD) and the viability determined by a CEDEX. B) Shown is the mean oxygen transfer rate (OTR). For clarity, only every 24th measuring point over time is marked as a symbol. The lines are drawn through all measured values. The outliners in the OTR data due to temperature adaptations after sampling were excluded from the data. In addition, the antibody concentration is shown. C) Corresponding glucose, glutamine, and lactate concentrations are plotted over time. Depletion of glucose and glutamine is marked by dotted vertical lines over all three parts of the figure. Cultivations were performed in a TOM device. Culture conditions: 250 mL TOM glass flasks, temperature (T) = 36.5 ◦C, shaking frequency (n) = 140 rpm, shaking diameter (d0) = 50 mm, filling volume (VL) = 50 mL, 5 % CO2, 70 % rel. hum., medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1.

Article Snippet: In this study, the suspension-adapted cell line CHO DP12 (clone#1934, ATCC CRL-12445) was utilized as a model production cell line.

Techniques: Sampling, Concentration Assay

Journal: New biotechnology

Article Title: Impact of hydromechanical stress on CHO cells' metabolism and productivity: Insights from shake flask cultivations with online monitoring of the respiration activity.

doi: 10.1016/j.nbt.2024.09.008

Figure Lengend Snippet: Fig. 2. Cultivation of CHO DP12 cells in shake flasks with varying average energy dissipation rates (εØ). The mean oxygen transfer rate (OTR) of three replicates is shown with standard deviations illustrated as shaded areas. For clarity, only every twelfth measuring point over time is marked as a symbol. The lines are drawn through all measured data points. The outliners in the OTR data due to temperature adaptations after opening the incubation hood were excluded from the data. The curves in dark and light red are from the inde pendent experiments depicted in Fig. 1 and Supp. Fig. S1. Culture conditions: flask size, shaking frequency, shaking diameter, and filling volume see Table 1, temperature (T) = 36.5 ◦C, 5 % CO2, 70 % rel. hum., medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1.

Article Snippet: In this study, the suspension-adapted cell line CHO DP12 (clone#1934, ATCC CRL-12445) was utilized as a model production cell line.

Techniques: Incubation

Journal: New biotechnology

Article Title: Impact of hydromechanical stress on CHO cells' metabolism and productivity: Insights from shake flask cultivations with online monitoring of the respiration activity.

doi: 10.1016/j.nbt.2024.09.008

Figure Lengend Snippet: Fig. 3. Correlation of the maximal specific growth rate (µmax) as well as the time of glucose and glutamine depletion to varying average energy dissipation rates (εØ) for CHO DP12 cell cultivations. εØ is plotted logarithmically. A) µmax calculated from the individual replicates of the OTR data in Fig. 2. B) Culti vation time difference to the standard cultivation with εØ = 0.12 W kg−1 of glutamine depletion read out from the glutamine shoulder in the OTR data in Fig. 2. C) Cultivation time difference to the standard cultivation with εØ = 0.12 W kg−1 of glucose depletion read out from the glucose drop in the OTR data in Fig. 2. A linear fit was performed for all figures (shown as a black line).

Article Snippet: In this study, the suspension-adapted cell line CHO DP12 (clone#1934, ATCC CRL-12445) was utilized as a model production cell line.

Techniques:

Journal: New biotechnology

Article Title: Impact of hydromechanical stress on CHO cells' metabolism and productivity: Insights from shake flask cultivations with online monitoring of the respiration activity.

doi: 10.1016/j.nbt.2024.09.008

Figure Lengend Snippet: Fig. 4. Analysis of the lactate concentration of CHO DP12 cell cultivations in shake flasks at varying average energy dissipation rates (εØ). A) Illustrated are the mean lactate concentrations of three biological replicates over time for the cultivations depicted in Fig. 2. B) The cultivation time difference to the standard cultivation with εØ = 0.12 W kg−1 of the lactate depletion read out from the offline data in A. εØ is plotted in logarithmic scale. Culture conditions: flask size, shaking frequency, shaking diameter, and filling volume see Table 1, temperature (T) = 36.5 ◦C, 5 % CO2, 70 % rel. hum., medium: TCX6D + 8 mM glutamine; starting cell density: 5 × 105 cells mL−1.

Article Snippet: In this study, the suspension-adapted cell line CHO DP12 (clone#1934, ATCC CRL-12445) was utilized as a model production cell line.

Techniques: Concentration Assay

Journal: New biotechnology

Article Title: Impact of hydromechanical stress on CHO cells' metabolism and productivity: Insights from shake flask cultivations with online monitoring of the respiration activity.

doi: 10.1016/j.nbt.2024.09.008

Figure Lengend Snippet: Fig. 5. Correlation of the final antibody concentration to the logarithm of varying average energy dissipation rates (εØ) for CHO DP12 cell cultivations. The mean value of the last three cultivation days was calculated. Statistically significant differences against the reference cultivation (εØ = 0.12 W kg−1) are indicated by stars (*** p < 0.001).

Article Snippet: In this study, the suspension-adapted cell line CHO DP12 (clone#1934, ATCC CRL-12445) was utilized as a model production cell line.

Techniques: Concentration Assay