Bioprocessing makes use of surface area such as packed beds and hollow fibers for scale up applications. In the same manner 3D porous scaffolds can serve the purpose to deliver both ample surface area and volumetric growth. Here, we present the 3D Perfusion Bioreactor.

1. Upstream
Processing:
Dimensionality
and
Perfusion
Add Extra Dimension to Your Innovation™
©
3D
BIOTEK,
LLC

2. 2D
Is
Easier,
at
What
Cost?
Scale-­‐up
ProducBon:
Cell
lines
and
Biologics
2D
Fermenter/2D
DissociaBon/Maintenance
of
Cells/Vaccine
ProducBon
1900’s
1960’s
2000’s
Glass-­‐Based
Tissue
Culture
2D
Polystyrene-­‐Based
Tissue
Culture
www.Pubmed.gov
2
2
©
3D
BIOTEK,
LLC

3. 1.
Lack
of
Dimensional
Relevance
Brain
Brain
Lung,
Breast
Lung,
Breast
Liver
Liver
≠
Bone,
CarDlage
Bone,
CarBlage
Petri
Dish
3
©
3D
BIOTEK,
LLC

4. 2.
Lack
of
3D
Perfusion
3D
Scaffold
2D
Surface
ConvecBve
Flow
≠
Perfusion
Flow
UpregulaBon
of
global
gene
expression
Uncorrelated
Gene
Expression
e.g.
ECM,
proliferaBon,
and
differenBaBon
DownregulaBon
of
hypoxia-­‐related
genes
4
©
3D
BIOTEK,
LLC

5. 3.
Lack
of
volumetric
Growth
3D
Scaffold
2D
Surface
Monolayer
Tissue-­‐Like
≠
Longer
ProliferaDon
Profiles
Shorter
ProliferaDon
Profiles
5
©
3D
BIOTEK,
LLC

6. 4.
Lack
of
Scalability
Total
Area
3D
35-­‐mm
T-­‐175
in
cm^2
Chambers
Flask
2,809.0
4
16
1
Loaded
Chamber
=
4
T175
Flasks
35-­‐mm
Chamber
6
©
3D
BIOTEK,
LLC

7. 3D
Technology:
Process-­‐Friendly
and
Biologically
Relevant
 Polymer
Scaffold
Prototyping
 3D
Perfusion
and
Seeding
OpDmizaDon
7
©
3D
BIOTEK,
LLC

8. 3D
Insert™
PlaWorm
Fiber
Diameter
Fiber-­‐to-­‐Fiber
Spacing
Scaffold
Layering
Pore
Size
Scaffold
Volume
3D
Geometry
8
©
3D
BIOTEK,
LLC

9. 3D
Insert™
Series
3D
Insert™-­‐Polystyrene

ConfiguraBons:
PS(1520)
and
PS(3040)
 PS(1520)
=
150-­‐um
Fiber/200-­‐um
Spacing
 PS(3040)
=
300-­‐um
Fiber/400-­‐um
Spacing
 Fit
96
to
6
well
plate
formats
 Total
number
of
layers:
4
3D
Insert™-­‐Poly-­‐ε-­‐Caprolactone

ConfiguraBons:
PCL(3030)
and
PS(3050)
 PCL(3030)
=
300-­‐um
Fiber/300-­‐um
Spacing
 PCL(3050)
=
300-­‐um
Fiber/500-­‐um
Spacing
 Fit
96
to
6
well
plate
formats
 Total
number
of
layers:
6
9
©
3D
BIOTEK,
LLC

10. 3D
Insert™:
Tissue-­‐like
Behavior
3D
Insert™
Fiber
Curvature
NIH
3T3
MCF-­‐7
Fiber
2
Fiber
1
H.
Astrocytes
hMSCs
Pore
Structure
10
©
3D
BIOTEK,
LLC

11. Ample
Surface
Area
in
Small
Volume
Average Total Cell Growth Area
2D 3D InsertTM-PS 3D InsertTM-PCL
6 well 6 well 6 well
1520 54.02 cm2 3030 99.21 cm2
9.6 cm2
3040 52.10 cm2 3050 75.62 cm2
12 well 12 well 12 well
1520 21.08 cm2 3030 39.27 cm2
4 cm2
3040 19.65 cm2 3050 27.90 cm2
24 well 24 well 24 well
1520 10.20 cm2 3030 18.28 cm2
1.9 cm2
3040 9.56 cm2 3050 13.74 cm2
48 well 48 well 48 well
1520 4.28 cm2 3030 7.74 cm2
1 cm2
3040 3.78 cm2 3050 6.08 cm2
96 well 96 well 96 well
1520 1.36 cm2 3030 2.03 cm2
0.32 cm2
3040 1.21 cm2 3050 1.53 cm2 11
©
3D
BIOTEK,
LLC

12. 3D
Technology:
Perfusion
12
©
3D
BIOTEK,
LLC

13. 3D
Perfusion:
Large-­‐Scale
Perfusion
3D
Chambers
Pump
Media
Reservoir
13
©
3D
BIOTEK,
LCC
CO2/Humidifier

14. Parts
and
Features
14
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3D
BIOTEK,
LLC

15. 3D
Perfusion
Bioreactor:
1.
Hardware
•
Durable
Metal
Frame

The
metal
frame
is
made
of
strengthened
aluminum
(anodized)
for
prolonged
durability
during
autoclaving.

Low
humidity
peristalBc
pump
with
maximum
60
rpm
15
©
3D
BIOTEK,
LLC

16. 3D
Perfusion
Bioreactor:
2.
Liquid
Handling
Media
Bo)les
Humidifier
Bo)les
•
Economic
and
Re-­‐usable

Media
and
humidified
boeles
and
accessories
are
autoclavable,
e.g.
glass
boeles,
stainless
steel,
silicon
gaskets,
and
polycarbonate
caps.
Total
Media
Capacity:
400-­‐ml
and
2-­‐liters
16
©
3D
BIOTEK,
LLC

17. 3D
Perfusion
Bioreactor:
3.
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
peristalBc
tubing
17
©
3D
BIOTEK,
LLC

18. Features:
1.
Color-­‐Coded
Assembly
O
Y
G
R
B
G
Media
Reservoir
Media
Reservoir
CO2/O2
Humidifie CO2/O2
Humidifier
r
18
©
3D
BIOTEK,
LLC

19. Features:
2.
ConDnuous
Perfusion
3D
Bioreactor
Perfusion
Circuitry
19
©
3D
BIOTEK,
LCC

20. Feature:
3.
Variable
Velocity
Range
Bioreactor
Chamber
Format
24-­‐Well
Insert
12-­‐Well
Insert
6-­‐Well
Insert
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
SpecificaDons)
20
©
3D
BIOTEK,
LLC

21. Features:
4.
Streamlined
Process
Media
Out
3
h
incubaDon
2.0
rpm,
24h
37º
C,
5%
CO2
Slow
Perfusion
For
24
well
format
Cell
Seeding
O-­‐Ring
3D
Insert™
Media
In
For
Perfusion
Bioreactor,
please
visit
us
at
hp://www.3dbiotek.com/web/prod_3dbioreactor.aspx
21
©
3D
BIOTEK,
LLC

22. 3D
Perfusion
Bioreactor
•
An
innovaBve
product
for
scale-­‐up
cell
culture
and
bioprocessing
•
The
only
polymer
scaffold-­‐based
perfusion
bioreactor
for
3D
dynamic
cell
culture
•
Simple
design
and
least
number
of
parts
for
faster
assembly
and
beeer
asepBc
condiBons
22
©
3D
BIOTEK,
LLC