Engineering Bioreactor Cell Engineering Biology Cell Stability Selection

Engineering Bioreactor Cell Engineering Biology Cell Stability & Selection Monitoring & Control Therapeutics & Diagnostics Tissue Engineering Medicine

Increasing Number of Cell Lines Being Maintained and Used More Cellular Assays Using New Markers and Dyes Cell Analysis Emphasis on Greater Throughput and Precision Pressure to Reduce Time, Cost and Complexity

CHONDROPROGENITOR CELLS CULTURED ON CULTISPHER-G Process outline PASSAGE PERFORMANCE Initial Cell Density (cells/ml) 3. 3 x 10 4 Final Cell Density (cell/ml) 4. 95 x 10 5 Microcarrier Concentration (mg/ml) 0. 6 Passage Length (h) Passage Population Doubling (-) Passage Expansion Factor (-) 120 3. 91 15 1500 L 1. 1 x 1013 cells 7 L 106 cells P 1 Inoculum (cells) Microcarriers (mg) Medium Volume (ml) Final Cell Number (cells) 1 x 106 18 30 1. 5 x 10 7 P 2 P 3 P 4 1. 5 x 10 7 2. 7 x 10 2 4. 5 x 10 2 2. 3 x 10 8 4. 1 x 10 3 6. 8 x 10 3 3. 4 x 10 9 6. 1 x 10 4 1 x 105 5. 1 x 10 10 P 5 5. 1 x 10 10 9. 1 x 10 5 1. 5 x 10 6 7. 6 x 10 11 P 6 7. 6 x 10 11 1. 4 x 10 7 2. 3 x 10 7 1. 1 x 10 13 30 days EXPANSION PERFORMANCE Monolayer Initial Cell Number (cells) Total Microcarriers (mg) Total Medium Volume (l) Final Cell Number (cells) 1 x 106 1. 5 x 10 7 2. 4 x 10 4 1. 1 x 10 13 Final Population Doubling (-) Total Expansion Factor (-) Total Expansion Time (days) Total Passage Number (-) 23. 44 1. 1 x 10 7 30 6 Initial Cell Number (cells) Total Area (cm 2) Total Medium Volume (ml) Final Cell Number (cells) 1 x 106 5. 4 x 10 8 1. 6 x 10 8 1. 1 x 10 14 Final Population Doubling (-) Total Expansion Factor (-) Total Expansion Time (days) Total Passage Number (-) 26. 76 1. 1 x 10 8 30 6

Chondrocyte culture into monolayer systems Isolated chondrocytes Expanded chondrocytes Biopsy removal Phase 1 CHONDROCYTE EXPANSION Cell bank for future uses Phase 2 in vitro FORMATION OF FUNCTIONAL TISSUE EQUIVALENTS Construct implantation Biocompatible-biodegradable scaffold Engineered cartilage Cell/scaffold constructs culture in tissue bioreactor