Journal: Bioactive Materials

Article Title: Upscaling human mesenchymal stromal cell production in a novel vertical-wheel bioreactor enhances extracellular vesicle secretion and cargo profile

doi: 10.1016/j.bioactmat.2022.07.004

Figure Lengend Snippet: hMSC expansion and metabolism on Synthemax II microcarriers in PBS Vertical-Wheel Bioreactors at 25, 40, and 64 RPM. (A) Process flow for hMSC expansion on Synthemax II microcarrers in PBS Vertical-Wheel Bioreactors; (B) hMSC growth kinetics during expansion and EV collection stages; (C) Glucose and Lactate metabolism: (i) Glucose concentration kinetics; (ii) Lactate concentration kinetics. (iii) Glucose consumed normalized to cell number; (iv) Lactate produced normalized to cell number; (v) Lactate production to glucose consumption ratios. (D) Glutamine metabolism: (i) Glutamine concentration kinetics. The extracellular vesicle (EV) collection medium is formulated with Glutamax. Therefore, the glutamine concentration at day 5 is 0. (ii) Glutamic acid concentration kinetics; (iii) Ammonia concentration kinetics.

Article Snippet: PBS 0.1L and 0.5L bioreactor systems were inoculated in parallel alongside Corning CellBIND T-flasks ( A).

Techniques: Concentration Assay, Produced

Journal: Bioactive Materials

Article Title: Upscaling human mesenchymal stromal cell production in a novel vertical-wheel bioreactor enhances extracellular vesicle secretion and cargo profile

doi: 10.1016/j.bioactmat.2022.07.004

Figure Lengend Snippet: Metabolic and extracellular vesicle (EV) biogenesis pathways and reactive oxygen species (ROS) analysis for hMSCs in PBS Vertical-Wheel Bioreactors at 25, 40, and 64 RPM. Various genes were measured by RT-qPCR on the mRNAs isolated from the harvested cells at the end of EV collection (n = 6, results were from two independent bioreactor runs), relative to 2D control. (A) Glycolytic genes; (B) Autophagy genes; Flow cytometry histograms for (C) Total ROS; (D) Mitochondrial ROS; and (E) Autophagosomes. Red line: negative control; blue line: 2D culture control; orange line: 25 RPM bioreactor culture; light green: 40 RPM, and dark green: 64 RPM. (F) ESCRT-dependent EV biogenesis markers; (G) ESCRT-independent EV biogenesis markers. * indicates p < 0.05; ** indicates p < 0.01, *** indicates p < 0.001.

Article Snippet: PBS 0.1L and 0.5L bioreactor systems were inoculated in parallel alongside Corning CellBIND T-flasks ( A).

Techniques: Quantitative RT-PCR, Isolation, Control, Flow Cytometry, Negative Control

Journal: Bioactive Materials

Article Title: Upscaling human mesenchymal stromal cell production in a novel vertical-wheel bioreactor enhances extracellular vesicle secretion and cargo profile

doi: 10.1016/j.bioactmat.2022.07.004

Figure Lengend Snippet: hMSC expansion and metabolism on Synthemax II microcarriers in 0.5L PBS Vertical-Wheel Bioreactors. (A) Process flow for hMSC expansion on Synthemax II microcarrers in PBS Vertical-Wheel Bioreactors; (B) hMSC numbers during expansion and EV collection stages; (C) Glucose and Lactate metabolism: (i) Glucose concentration kinetics; (ii) Lactate concentration kinetics. (iii) Glucose consumed normalized to cell number; (iv) Lactate produced normalized to cell number; (v) Lactate production to glucose consumption ratios. (D) Glutamine metabolism: (i) Glutamine concentration kinetics. The EV collection medium is formulated with Glutamax. (ii) Glutamic acid concentration kinetics; (iii) Ammonia concentration kinetics.

Article Snippet: PBS 0.1L and 0.5L bioreactor systems were inoculated in parallel alongside Corning CellBIND T-flasks ( A).

Techniques: Concentration Assay, Produced

Journal: Bioactive Materials

Article Title: Upscaling human mesenchymal stromal cell production in a novel vertical-wheel bioreactor enhances extracellular vesicle secretion and cargo profile

doi: 10.1016/j.bioactmat.2022.07.004

Figure Lengend Snippet: Metabolic and extracellular vesicle biogenesis pathways for hMSCs in 0.5L PBS Vertical-Wheel Bioreactors. Various genes were measured by RT-qPCR on the mRNAs isolated from the harvested cells at the end of EV collection (n = 6, results were from two independent bioreactor runs), relative to 0.1L bioreactor control. (A) Glycolytic genes; (B) Autophagy genes; (C) ESCRT-dependent EV biogenesis markers; (D) ESCRT-independent EV biogenesis markers. * indicates p < 0.05; ** indicates p < 0.01, *** indicates p < 0.001.

Article Snippet: PBS 0.1L and 0.5L bioreactor systems were inoculated in parallel alongside Corning CellBIND T-flasks ( A).

Techniques: Quantitative RT-PCR, Isolation, Control