Journal: Bioengineering

Article Title: Bioreactor Parameters for Microcarrier-Based Human MSC Expansion under Xeno-Free Conditions in a Vertical-Wheel System

doi: 10.3390/bioengineering7030073

Figure Lengend Snippet: Fed-batch bioreactor culture paradigm. ( A ) Comparison study of a xeno-free (XF) bioreactor process utilizing batch and fed-batch processes shows a distinct advantage of the fed-batch process on final cell yield ( n = 3). ( B ) Media productivity is enhanced using a fed-batch process. ( C ) Images indicating typical cell growth on microcarriers during a 5-day culture in a vertical-wheel bioreactor (* p ≤ 0.05). ( D ) Schematic of the bioreactor culture workflow using a fed-batch process.

Article Snippet: Following the seed train, hMSCs were cultured on Corning ® Low Concentration SynthemaxTM II microcarriers (Corning) inside PBS-0.1 single-use Vertical-WheelTM bioreactors (PBS Biotech, Camarillo, CA, USA).

Techniques: Comparison

Journal: Bioengineering

Article Title: Bioreactor Parameters for Microcarrier-Based Human MSC Expansion under Xeno-Free Conditions in a Vertical-Wheel System

doi: 10.3390/bioengineering7030073

Figure Lengend Snippet: Determination of cell and microcarrier inoculation number. ( A ) Growth kinetics in bioreactors for various cell numbers and microcarrier during seeding, as well as media exchanges. Cell counts were obtained from sampling. ( B ) Final harvest density obtained from in-vessel harvest. At the highest tested cell-to-media ratio, cell growth can only be supported through media exchanges. ( C ) Media productivity comparison shows the disadvantage of media exchanges despite the high cell yield. Tested groups are 1×: 2.1 × 10 6 cells and 1.25 g; 2×: 4.2 × 10 6 cells and 2.5 g; 3×: 6.3 × 10 6 cells and 3.75 g; and 3×ΔΔ: 6.3 × 10 6 cells and 3.75 g with media exchanges on Days 3 and 4.

Article Snippet: Following the seed train, hMSCs were cultured on Corning ® Low Concentration SynthemaxTM II microcarriers (Corning) inside PBS-0.1 single-use Vertical-WheelTM bioreactors (PBS Biotech, Camarillo, CA, USA).

Techniques: Sampling, Comparison

Journal: Bioengineering

Article Title: Bioreactor Parameters for Microcarrier-Based Human MSC Expansion under Xeno-Free Conditions in a Vertical-Wheel System

doi: 10.3390/bioengineering7030073

Figure Lengend Snippet: Optimization of bioreactor seeding parameters at fixed microcarrier amount (A; 1.25 g), or fixed cell number (B; 2.1 × 10 6 cells). ( A ) Effect of cell seeding numbers on bioreactor culture yield with 1.25 g microcarriers, and ( B ) effect of microcarrier density on bioreactor culture yield with 2.1 × 10 6 cells seeded. For both optimizations, we presented hMSC growth kinetics in bioreactors based on cell density from daily sampling (i), final density obtained from an in-vessel harvest (ii), expansion of cells harvested from the bioreactors compared to the 2D control (iii), and analysis of nutrient and waste in medium during culture (iv). Both seeding parameter studies support bioreactor seeding with 1.25 g of microcarriers and 2.1 × 10 6 cells (4667 cells/cm 2 ) to optimize for the cell yield and post-bioreactor expansion, while minimizing costs associated with materials.

Article Snippet: Following the seed train, hMSCs were cultured on Corning ® Low Concentration SynthemaxTM II microcarriers (Corning) inside PBS-0.1 single-use Vertical-WheelTM bioreactors (PBS Biotech, Camarillo, CA, USA).

Techniques: Sampling, Control

Journal: Bioengineering

Article Title: Bioreactor Parameters for Microcarrier-Based Human MSC Expansion under Xeno-Free Conditions in a Vertical-Wheel System

doi: 10.3390/bioengineering7030073

Figure Lengend Snippet: Optimization of bioreactor culture process. ( A ) Effect of bioreactor agitation speed on hMSC growth. Higher agitation speed (45 rpm) results in lower final cell yield. ( B ) Cell health following bioreactor culture. Cells harvested from the bioreactor on Day 6 showed 30% reduced expansion potential. ( C , D ) Effect of bioreactor feed timing on hMSC growth. Addition of feed on Day 3 results in the highest cell yield. ( E ) Effect of microcarrier addition during culture. Addition of 1.25 g of microcarriers (additional 450 cm 2 of surface area) to increase growth surface area during culture did not influence overall hMSC growth.

Article Snippet: Following the seed train, hMSCs were cultured on Corning ® Low Concentration SynthemaxTM II microcarriers (Corning) inside PBS-0.1 single-use Vertical-WheelTM bioreactors (PBS Biotech, Camarillo, CA, USA).

Techniques:

Journal: Bioengineering

Article Title: Bioreactor Parameters for Microcarrier-Based Human MSC Expansion under Xeno-Free Conditions in a Vertical-Wheel System

doi: 10.3390/bioengineering7030073

Figure Lengend Snippet: Effect of surfactants on hMSC growth in bioreactor. Addition of pluronic PF68 shows no effect on cell growth ( A ), however a high concentration of PF68 (2 g/L) markedly reduces post-bioreactor expansion ( B ).

Article Snippet: Following the seed train, hMSCs were cultured on Corning ® Low Concentration SynthemaxTM II microcarriers (Corning) inside PBS-0.1 single-use Vertical-WheelTM bioreactors (PBS Biotech, Camarillo, CA, USA).

Techniques: Concentration Assay

Journal: Bioengineering

Article Title: Bioreactor Parameters for Microcarrier-Based Human MSC Expansion under Xeno-Free Conditions in a Vertical-Wheel System

doi: 10.3390/bioengineering7030073

Figure Lengend Snippet: Effect of various bioreactor harvest conditions on post-harvest cell health: ( A ) agitation duration and speed during harvest in TrypLE, ( B ) duration in quench solution, and ( C ) temperature of quench solution.

Article Snippet: Following the seed train, hMSCs were cultured on Corning ® Low Concentration SynthemaxTM II microcarriers (Corning) inside PBS-0.1 single-use Vertical-WheelTM bioreactors (PBS Biotech, Camarillo, CA, USA).

Techniques:

Journal: Bioengineering

Article Title: Bioreactor Parameters for Microcarrier-Based Human MSC Expansion under Xeno-Free Conditions in a Vertical-Wheel System

doi: 10.3390/bioengineering7030073

Figure Lengend Snippet: Validation of optimized fed-batch bioreactor process and hMSC characterization following bioreactor culture. ( A ) Parameters for fed-batch culture in a bioreactor. ( B ) Validation of optimized bioreactor culture process in five hMSC donors. ( C – G ) Verification of cell health, critical quality attributes, and functionality following bioreactor culture: XF hMSCs expanded in the bioreactor maintained their cell surface marker expression identity ( C ), tri-lineage differentiation potential to osteo-, adipo-, and chondrocytes ( E ), inducible indoleamine 2,3-dioxygenase (IDO) activity when stimulated with IFN γ ( F ), and cytokine secretion profile ( G ). The health, critical quality attributes, and functionality of three hMSC donors are comparable for cells harvested from bioreactor and cells harvested from its respective 2D flask control (In panel E, Scale bar = 400 µm).

Article Snippet: Following the seed train, hMSCs were cultured on Corning ® Low Concentration SynthemaxTM II microcarriers (Corning) inside PBS-0.1 single-use Vertical-WheelTM bioreactors (PBS Biotech, Camarillo, CA, USA).

Techniques: Biomarker Discovery, Marker, Expressing, Activity Assay, Control