3D Perfusion Bioreactor Technical Presentation
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The 3D Perfusion Bioreactor™ is manufactured by 3D Biotek, LLC, an innovative technology company based in North Brunwick, New Jersey. 3D Biotek launched the 3D Perfusion Bioreactor™ in 2011, as a novel alternative to integrate the need for innovative cellular maintenance and increasing trends towards 3-dimensional cell culture. The 3D Perfusion Bioreactor is engineered to perfuse porous 3D polymer Inserts™ in 4 separate polycarbonate chambers able to fit up to 10-scaffolds per chamber. The 3D Insert has been validated by NIST (National Institute of Standards) for its open-pore, ample surface area, and 100% interconnected geometry, important features for direct application in scale-up processes, from cell expansion to recombinant protein production.
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3D Perfusion Bioreactor Technical Presentation
- 1. 3D Perfusion Bioreactor™ Add Extra Dimension to Your Innovation 1 © 3D BIOTEK, LLC
- 2. Agenda/Learning Objec0ves • Introduc*on • Product Descrip*on • Opera*on Principle • Applica*ons 2 © 3D BIOTEK, LLC
- 3. Introduc>on 3 © 3D BIOTEK, LLC
- 4. 3D Perfusion Bioreactor™ • An innova*ve product for cell culture and bioprocessing • The only polymer scaffold-‐based perfusion bioreactor for 3D dynamic cell culture • Simple design and minimal parts for faster assembly and beAer asep*c condi*ons 4 © 3D BIOTEK, LLC
- 5. 3D Perfusion Bioreactor Adds Value • Proven 3D InsertTM technology, scaffolds with 100% open pore structure and pore Interconnec*on • Choice of both biodegradable and non-‐ biodegradable scaffolds. 5 © 3D BIOTEK, LLC
- 6. 3D Perfusion Bioreactor Adds Value, Cont • Process can be standardized, scaffolds were selected as Reference scaffold by NIST • Adjustable dynamic perfusion velocity range • BoAom to top flow to maintain even wePng of stacked scaffolds 6 © 3D BIOTEK, LLC
- 7. Fields of Applica0on • Stem cell/*ssue engineering Bone, car*lage, liver, etc. • In vitro *ssue model for drug discovery Tumor and liver models • Bioprocessing Cell expansion (enhanced prolifera*ve capacity) Recombinant protein Hybridoma-‐based an*body produc*on 7 © 3D BIOTEK, LLC
- 8. Product Descrip>on 8 © 3D BIOTEK, LLC
- 9. Product Specifica0on • Autoclavable System (Except Pump) • Low Humidity Working Pump • Color Coded Tubing for Easy Assembly • Op*mized Cell Seeding in Scaffolds • Adjustable Perfusion Veloci*es • 3D Inserts™ wide range of pore sizes Any polymer of interest 9 © 3D BIOTEK, LLC
- 10. 3D Perfusion Bioreactor™ Diagram 5 1 4 4 1 9 2 1 9 3 1 8 1 3 1 7 1 0 1 Z 5 Y 2 X 10 © 3D BIOTEK, LLC
- 11. 3D Perfusion Bioreactor: Hardware • Durable Metal Frame The metal frame is made of strengthened aluminum (anodized) for prolonged durability during autoclaving. Low humidity peristal*c pump with maximum 60 rpm. 11 © 3D BIOTEK, LLC
- 12. 3D Perfusion Bioreactor™: Perfusion Circuit 12 © 3D BIOTEK, LLC
- 13. 3D Perfusion Bioreactor: Liquid Handling Media BoJles • Economic and Re-‐usable Media and humidified boAles and accessories are autoclavable, e.g. glass boAles, stainless steel, silicon gaskets, and polycarbonate caps. 13 © 3D BIOTEK, LLC
- 14. 3D Perfusion Bioreactor: Tubing System Tubing and Transport (A) Perfusion Chamber (B) Media System (C) CO2/O2 System (A) (C) (B) • Media and Gas Transport Medical grade clear silicon and beige peristal*c tubing 14 © 3D BIOTEK, LLC
- 15. 3D Perfusion Bioreactor: Color -‐coded Assembly O Y G R B G Media Reservoir Media Reservoir CO2/O2 Humidifie CO2/O2 Humidifier r 15 © 3D BIOTEK, LLC
- 16. 3D Insert™ Scaffold Geometry 3D Insert™-‐Polystyrene Polystyrene (PS) is a transparent polymer used in tradi*onal cell culture. 3D Insert™-‐Poly-‐ε-‐Caprolactone Poly-‐ε-‐Caprolactone (PCL) is a biodegradable polymer used in FDA approved medical devices 16 © 3D BIOTEK, LLC
- 17. 3D Insert™ Scaffold Specifica0ons 3D Insert™-‐Polystyrene Configura*ons: PS(1520) and PS(3040) PS(1520) = 150-‐um Fiber/200-‐um Spacing PS(3040) = 300-‐um Fiber/400-‐um Spacing Total number of layers: 4 3D Insert™-‐Poly-‐ε-‐Caprolactone Configura*ons: PCL(3030) and PS(3050) PCL(3030) = 300-‐um Fiber/300-‐um Spacing PCL(3050) = 300-‐um Fiber/500-‐um Spacing Total number of layers: 6 17 © 3D BIOTEK, LLC
- 18. 3D Insert™ Technology Fiber Diameter Fiber-‐to-‐Fiber Spacing Scaffold Layering Pore Size Scaffold Volume 3D Geometry 18 © 3D BIOTEK, LLC
- 19. 3D Perfusion: Specific Product Features • Easy –Handle 3D Insert™ Scaffolds adapted for beAer handling Increase loading precision Faster loading *me • Chamber Lateral Micro Channels Polycarbonate chamber 0.6-‐mm channels inside the chamber Prevent bubbling during ini*al media flow Helps maintain op*mum ini*al seeding density 19 © 3D BIOTEK, LLC
- 20. 3D Insert™: Ideal Scaffold for Perfusion Bioreactor Matrigel / PuraMatrix / AlgiMatrix 3D Calcium Phosphate 3D Collagen 3D OPLA Alvatex® Coa>ngs Scaffold Scaffold Scaffold High Variable 100% Transparency (direct Ready surface to configurations Easy cell Plate reader interconnected observation with to use volume recovery compatible pores (customizable) light microscope) ratio 3D Insert™ Gel Matrices PLA foam CaP foam Alginate Foam Alvetex® Compa>ble Not Compa>ble 20 © 3D BIOTEK, LLC
- 21. Product Comparison: Bioreactors Easy High Variable 100% Cell surface to configurations Continuou 3D Dynamic interconnect Recover volume s Shear Perfusion ed pores (customizable) y ratio 3D Perfusion Roller Bottle Fiber Cell Stirrer Flask BioLevitaror Compa>ble Not Compa>ble 21 © 3D BIOTEK, LLC
- 22. Opera>on Principle 22 © 3D BIOTEK, LLC
- 23. 3D Perfusion Bioreactor™: Cell Seeding Media Out 3 h incuba*on 1.93 rpm, 24h 37º C, 5% CO2 Slow Perfusion Cell Seeding O-‐Ring 3D Insert™ Media In © 3D BIOTEK, LLC 3D Seeding 3D Perfusion 23
- 24. 3D Perfusion: Cell Seeding Volumes Seeding Volume (ul) Insert™ PS(1520) PS(3040) PS(3030) PS(3050) Format 24-‐well 60 120 130 160 12-‐well 120 240 270 340 6-‐well 300 600 690 880 24 © 3D BIOTEK, LLC
- 25. 3D Perfusion: Cell Culture Media Volumes Bioreactor Format and Volume to Fill (ml) Tubing 6 –Well Chamber 45-‐ml 12-‐Well Chamber 15-‐ml 24-‐Well Chamber 7.0-‐ml Media Tubing 5.0-‐ml Reservoir BoEle Max. volume 90-‐ml 25 © 3D BIOTEK, LLC
- 26. 3D Perfusion: rpm and Velocity Bioreactor Chamber Format 24-‐Well 12-‐Well 6-‐Well Velocity(cm/min) rpm rpm rpm 0.0 0 0 0 0.2 1.93 4.84 12.08 0.4 3.86 9.69 24.15 0.6 5.8 14.53 36.23 0.8 7.73 19.38 48.31 *Pump maximum rpm capacity is 60 rpm (Manufacturer Specifica*ons) 26 © 3D BIOTEK, LLC
- 27. Applica>ons 27 © 3D BIOTEK, LLC
- 28. 3D Perfusion: Increased Cell Viability 6 5 2 Cell Fold Increase Top 4 3 4 2 1 0 6 1 3 5 7 9 Scaffold Posi>on (Top (Lec) to Bodom (Right)) 8 BoAom 10 Dynamic Sta>c Neonatal Fibroblats Calcein Viability Neonatal Fibroblats Growth in Perfusion 28 © 3D BIOTEK, LCC
- 29. 3D Perfusion: Superior Cell Prolifera0on Day 5 n = 3 (STDEV) 3D Perfusion Fiber 300-‐um 3D Sta>c 3D Perfusion Causes Early Prolifera0on of Osteosarcoma 29 © 3D BIOTEK, LCC
- 30. 3D Perfusion: Enhances in vivo-‐like Behavior Total Collagen Staining Shows Increasing Deposi0on of ECM 30 © 3D BIOTEK, LCC
- 31. Visit us @ hEp://www.3dbiotek.com/web/ 31 © 3D BIOTEK, LLC