Product Description

LifeSupport® is the core component and industry standard for FRESH bioprinting.

FRESH stands for Freeform Reversible Embedding of Suspended Hydrogels.

Each LifeSupport® printing kit comes with 4 units of 2 grams of sterile LifeSupport® powder. One gram of powder hydrates to approximately 10 mL of FRESH support material.

Read the groundbreaking publication using LifeSupport® here: https://science.sciencemag.org/content/365/6452/482

Parameter, Testing, and Method LifeSupport® #5244
Sterilization Method Irradiation
Sterility - USP modified No growth
Form Grounded Powder
Bioprinting Supports Lifeink® Collagen Bioprinting
Average Particle Size Diameter 15-50 microns
Package Size 8 grams of powder (4 x 2 grams)
Rheology Bingham Plastic Properties
Shelf Life Minimum of 6 months from date of receipt
Source Material Bovine Gelatin

Directions for Use

Download the full PDF version or continue reading below:

1. Transfer the required weight of LifeSupport powder into a 50 mL sterile conical centrifuge tube (see Table 2 for recommended weights). Do not weigh out more than 2.1 grams per centrifuge tube.

2. Weigh out required powder into a 50 mL centrifuge tube, and then add recommended volume of COLD media as found in Table 2. Note – these volumes are to fill the printing dish, which is not needed for all prints. For example, printing a flat scaffold may only require the dish to be partially filled.

 

Table 2:

Printing Dish

Powder Weight

Media Volume

35 mm dish

1.0 gram

17 mL

6-well plate

2.10 grams

35 mL

12-well plate

0.86 grams

15 mL

24-well plate

0.43 grams

7 mL

 

LifeSupport™ can be customized to fit your specific bioink.
See the Table 3 below for help on choosing the right suspension media to take advantage of the unique benefits of FRESH bioprinting.

Table 3:

Bioink Suspension
Media
Centrifugation
Neutralized Collagen Cell Culture Media or 1X PBS 2000G*; 5 min
Alginate 0.1 wt% CaCl2 2000G; 5 min
Fibrinogen Media with Thrombin 2500G*; 5 min
Matrigel Cell Culture Media or 1X PBS 2000G*; 5 min
Acidic Collagen 50 mM HEPES buffer (pH 7.4) 2000G; 5 min
Decellularized ECM Cell Culture Media or 1X PBS 2000G*; 5 min

 

*Note: centrifugation speed and time may vary slightly depending on the suspension media. Serum-based growth media requires higher centrifugation speed (ie. 2500-3000G)

3. Vortex for 1 minute, ensuring that all the powder is fully resuspended and then allow to sit for 10 minutes while the powder fully rehydrates.

4. Centrifuge the support material according to table 3.

5. Support material should be compacted at the bottom of the tube. Pour off the supernatant.

6. Aseptically scoop out the compacted LifeSupport slurry into the desired printing dish using a sterile spatula or other device. Avoid introducing large air bubbles in the slurry. The slurry should not move if the dish is held sideways.

You are ready for FRESH bioprinting.

Printing Recommendations:
1. Place LifeSupport support slurry in your 3D bioprinter.

2. Calibrate (zero) your needle near the bottom of the dish (z), and near the middle (x,y). See below.

Note: Unlike typical printing, the needle does not have to be touching or even close to the bottom of the dish. The LifeSupport support slurry will trap your print in-place, no matter where you start. Ensure that the print container is large enough to avoid the needle hitting into the walls during printing.

3. Bioprinting can now be initiated.  Ensure the container will not shift on the print bed.
Note – It is possible to print multiple constructs in the same container in order to maximize the number of prints per LifeSupport kit.

Post Print Recommendations:
1. After printing, incubate the printed construct at 37°C for at least 30 minutes to allow the LifeSupport slurry to melt and release the bioprinted construct.

2. Cellularized bioinks printed into cell media-based support material can be directly placed in a cell incubator for 24 hours before needing to exchange media (ensuring proper levels of CO2).

3. After 30 minutes of post-print incubation, the LifeSupport slurry will be fully melted and your printed structure will be released. Large volumes may require longer times to fully melt.

4. Transfer of bioprinted construct:

4a. Carefully transfer bioprinted constructs into desired cell culture medium.

4b. As an alternative, melted LifeSupport slurry can be serially replaced with  pre-warmed cell culture medium, to avoid handling the printed construct. For example, if you printed into a 6-well plate, this can be done by carefully aspirating out 2 mL of melted LifeSupport and adding 2 mL of pre-warmed cell culture medium. Repeat this process until all of the melted LifeSupport slurry has been replaced by medium.

5. Continue standard media changes in accordance with cell culture protocol.

LifeSupport is manufactured by FluidForm under license from Carnegie Mellon University Patent No. 10,150,258.

Product Bioprints

Full Scale Human Neonatal Heart

Printed in LifeSupport

Heart Valve - Closed

Printed in LifeSupport

Heart Valve - Open

Printed in LifeSupport

Meniscus

Printed in LifeSupport

Product References

References for FRESH Printing:

Lee, A. et al. 3D bioprinting of collagen to rebuild components of the human heart. Science 365, 482–487 (2019).

Thomas, H. & Adam, F. 3D printing complex scaffolds using freeform reversible embedding of suspended hydrogels (FRESH). Frontiers in Bioengineering and Biotechnology4,(2016).

Isaacson, A., Swioklo, S. & Connon, C. J. 3D bioprinting of a corneal stroma equivalent. Experimental Eye Research173,188–193 (2018).

Jeon, O., Lee, Y., Hinton, T., Feinberg, A. & Alsberg, E. Cryopreserved cell-laden alginate microgel bioink for 3D bioprinting of living tissues. Materials Today Chemistry12,61–70 (2019).

Maxson, Eva L., et al. "In vivo remodeling of a 3D-Bioprinted tissue engineered heart valve scaffold." Bioprinting (2019): e00059.

Product Certificate of Analysis

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Safety and Documentation

Safety Data Sheet

Certificate of Origin

Product Disclaimer

This product is for R&D use only and is not intended for human or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.