SLIDE 1
1
2
- What is the CellJet?
- Basic Functioning
- Live Cell Printing
- How did we use it?
- Validation as a tool for Biofabrication
- Future Directions
A Novel Technology for Bio-Printing Presented By: Brian Syverud - - PowerPoint PPT Presentation
A Novel Technology for Bio-Printing Presented By: Brian Syverud - - PowerPoint PPT Presentation
A Novel Technology for Bio-Printing Presented By: Brian Syverud Chirantan Kanani info@digilabglobal.com http://digilabglobal.com 1 Overview What is the CellJet? Basic Functioning Live Cell Printing How did we use it?
SLIDE 2
SLIDE 3
3
Basic Operation
SLIDE 4
4
System Capabilities
- Fast: Fills a 1536 well plate in <1 minute
- Flexible
- Multi-Channel: Separate paths for cells,
media, hyrdrogels, etc.
- 20 nl – 4 µl volume range
- Accurate: 10 µm spatial accuracy
- Gentle: Non-contact fluid path avoids cell
constriction
SLIDE 5
5
Bio-Printing Validation
- Printing Cells
- Viability
- Functionality
- Concentration consistency
- Printing Hydrogels
- Drop dispense at high viscosity
- 2D Shapes
- 3D Layering
- 3D Construct – cells in hydrogel architecture
SLIDE 6
6
Cell Viability
Human Muscle Stem Cells stained for live/dead assay
Scale bar: 200 μm
SLIDE 7
7
Cell Viability
20 40 60 80 100
Droplet Volume Cell Viability (%)
1 million cells/ml 2 million cells/ml
50nl 100nl 250nl 500nl
A
Smooth Muscle Cell Viability Human Muscle Stem Cell Viability
0.0 20.0 40.0 60.0 80.0 100.0 50 100 250 500
Droplet Volume (nl) Cell Viability (%)
1 million cells/ml
SLIDE 8
8
Cell Functionality
F-actin staining of Smooth Muscle Cells at 10x Magnification F-actin staining of Smooth Muscle Cells at 20x Magnification
SLIDE 9
9
Cell Consistency – Fluorescent Dye
Rhodamine fluorescence intensity as a function of Drop volume after CellJet Printing.
0.0 20.0 40.0 60.0 80.0 100.0 200 400 600 800 1000 1200 1400
Drop Volume (nl) Normalized Intensity
Average Value
SLIDE 10
10
Cell Consistency – Rat Arterial Smooth Muscle Cells
Cell concentration as a function of Drop volume after CellJet Printing
500 1000 1500 2000 1000 2000 3000
Drop Volume (nl)
Cell Count Expected Value
SLIDE 11
11
Cell Printing Validation
- Viability
- Smooth Muscle Cells
97.5%
- Human Muscle Stem Cells
98.4%
- Functionality
- F-actin Staining
- Consistency
- Fluorescent Dye Printing
± 4.8%
- Rat Arterial Smooth Muscle Cells
SLIDE 12
12
Hydrogel Printing
- Sodium Alginate
- Chemically dependent hydrogel – solidifies with Ca2+ ions
- Drop Dispense at High Viscosities
- 2D Shapes
- Lines
- Circles
- 3D Layering
SLIDE 13
13
Hydrogels – Drop Dispense
- 0.5% w/w Sodium Alginate
- Viscosity: 58.5 cP
- Volume Range: 100nl - 4µl
- 1.0% w/w Sodium Alginate
- Viscosity: 269.5 cP
- Volume Range: 125nl - 4µl
- 1.5% w/w Sodium Alginate
- Viscosity: 1180.2 cP
- Volume Range: 250nl - 4µl
SLIDE 14
14
Hydrogels – 2D Shapes
- Lines:
Average Line Thickness
0.00 0.10 0.20 0.30 0.40 0.50 1 2 3 4 5 6 7 8 9 10
Trial Thickness (mm)
Sodium Alginate gel printed to a glass slide
Mean: .357 ± .022mm
SLIDE 15
15
Hydrogels – 2D Shapes
- Circles:
Sodium Alginate printed in 10mm circle
Printed Geometry – controlled by Dispense Volume, Height, and Speed
Average Ring Thickness
0.00 0.50 1.00 1.50 1 2 3 4 5 6 7 8 9 10
Trial Thickness (mm)
Mean: 1.363 ± 0.129mm
SLIDE 16
16
Hydrogels – 3D Layering
- Sodium Alginate
- 2D Shapes – Controllable
- 3D Layering – Unexpected
Results
- Further Investigation
- Alternative Dispense Methods
- Other Hydrogels
SLIDE 17
17
Future Direction
- Print Hydrogels for support
- Print Cells suspended in Hydrogel in precise
locations
SLIDE 18
18
Bio-Printing Validation
- Printing Cells
- Viability > 95%
- Functionality
- Concentration consistency
- Printing Hydrogels
In Progress
- Drop dispense at high viscosity
- 2D Shapes
- 3D Layering
In Progress
- Future Direction – Functional 3D
Construct with Cells
SLIDE 19
19
- Sample. Science. Solutions.