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. 1. 3D  Perfusion  Bioreactor™     Add Extra Dimension to Your Innovation 1  ©  3D  BIOTEK,  LLC  
  2. 2. Agenda/Learning  Objec0ves   •   Introduc*on     • Product  Descrip*on   • Opera*on  Principle   • Applica*ons     2  ©  3D  BIOTEK,  LLC  
  3. 3. Introduc>on   3  ©  3D  BIOTEK,  LLC  
  4. 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. 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. 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. 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. 8. Product  Descrip>on   8  ©  3D  BIOTEK,  LLC  
  9. 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. 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. 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. 12. 3D  Perfusion  Bioreactor™:  Perfusion  Circuit         12  ©  3D  BIOTEK,  LLC  
  13. 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. 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. 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. 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. 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. 18. 3D  Insert™  Technology   Fiber  Diameter     Fiber-­‐to-­‐Fiber  Spacing      Scaffold  Layering         Pore  Size   Scaffold  Volume   3D  Geometry   18  ©  3D  BIOTEK,  LLC  
  19. 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. 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. 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. 22. Opera>on  Principle   22  ©  3D  BIOTEK,  LLC  
  23. 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. 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. 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. 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. 27. Applica>ons   27  ©  3D  BIOTEK,  LLC  
  28. 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. 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. 30. 3D  Perfusion:  Enhances  in  vivo-­‐like  Behavior       Total  Collagen  Staining  Shows  Increasing  Deposi0on  of  ECM   30  ©  3D  BIOTEK,  LCC  
  31. 31. Visit  us  @     hEp://www.3dbiotek.com/web/   31  ©  3D  BIOTEK,  LLC  

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