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3D Perfusion Bioreactor Technical Presentation
The 3D Perfusion Bioreactor™ is an innovative tool for dynamic cell culture and bioprocessing, utilizing a simple design with minimal parts for efficiency. It features proven 3D insert technology, adjustable perfusion velocities, and offers applications in tissue engineering, drug discovery, and bioprocessing. The bioreactor supports various scaffold types and is designed for enhanced cell viability and proliferation.
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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