Ref No: REP-VLA000001000
Date: 03-JAN-2023
Author: MG, AB, VK. Version: 2

RECONSTITUTION PROTOCOL

Pluronics PowderThis is a suggested procedure, please adjust according to your experimental needs.
To maintain the sterility of the product, work under sterile conditions.

Protocol aim

Materials needed

The aim of this protocol is to provide instructions forreconstituting Pluronics Powder. Pluronics is a sterile
powder that may be dissolved in water or a buffer
solution of your choice. Dissolved Pluronics exhibitshigh viscosity at room temperature and becomesliquid when cooled.

••••••••••Pluronics Powder*
Reconstitution liquid (e.g. water, PBS, HBSS,
cell culture medium etc.)
Tubes
Ice bath (for cooling)
Spatulas/spoons
Magnetic stirring plate
Magnetic stir bar
Cartridges, 3cc*
BIO X*, BIO X6* or INKREDIBLE series* 3D
bioprinterBioprinting nozzles* or needles**The product can be purchased in the CELLINKshop at www.cellink.com/shop.

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Protocol1.

Calculations

DESCRIPTION•••

Note the desired final concentration of Pluronics (cP). To achieve a gel that is 3D printable at room
temperature, the recommended final concentration is 40% w/v, cP.= 40.
Record the desired volume of gel to prepare, in mL (VL).
See Figure 1 for difference in viscosity of Pluronics gels of different concentrations.

•Calculate the amount of Pluronics powder to be used: 𝑚𝑃 = 𝐿 𝑃•

See Table 1 with calculations for suggested cP.

𝑉 ·𝑐
10Note: This equation gives a final concentration in weight/volume. However, depending on the concentrationof Pluronics, the gel may swell, resulting in slightly increased final volume.

Figure 1. Viscosity of Pluronics dissolved in deionized water at 20% and 40% over a shear rate range of 0.01 to100 s-1, 25°C.
Table 1. Suggested concentrations and the corresponding amount of Pluronics powder used for the preparation of
3 mL Pluronics gel.
Concentration of Pluronics gel, cP(%)
20%40%

2.

Volume of reconstitution liquid,
VL (mL)33

Mass of Pluronics powder, mP (g)0.6
1.2

Preparation of reconstitution solution

MATERIAL
Reconstitution liquidTubeDESCRIPTION••

Transfer VL of the reconstitution liquid to a centrifuge tube or container of choice.
Cool your reconstitution liquid to ~4°C using an ice bath or fridge.

Note: Since dissolved Pluronics becomes liquid at low temperature, while being a gel at room temperature,
the dissolution must be performed at low temperatures when making highly concentrated Pluronics gels. See
Figure 2 for temperature dependence of 20% and 40% Pluronics.

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Figure 2. Storage modulus of Pluronics dissolved in deionized water at 20% and 40% concentration, over atemperature range of 5-37°C.

3.

Preparation of Pluronics

MATERIAL
Spatula/spoon
Tube
Pluronics Powder

DESCRIPTION•4.

In another tube, weigh up mP of Pluronics Powder using a spatula/spoon.

Pluronics dissolution

MATERIAL
Cooled reconstitution liquid
Pluronics Powder
Magnetic stir bar

DESCRIPTION••••

Into the tube with reconstitution liquid, add the Pluronics Powder. To reduce the formation of clumps,
add the powder in increments and mix with a spatula.
Add a magnetic stir bar to the container.
Stir the mixture at high speed in an ice bath until dissolved. If clumps form, crush them with a spatula or
shake the container.
Store the Pluronics gel at 4-8°C.

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5.

3D printing

MATERIALPluronics gel
BIO X, BIO X6 or INKREDIBLE series 3D bioprinterBioprinting nozzle or needle

DESCRIPTION••

Transfer the Pluronics gel to a cartridge and cap with a bioprinting nozzle or needle.
3D print according to application. For suggestions on 3D printing using 40% Pluronics gel as a sacrificial
ink, see the Printing Protocol Pluronics 40%.

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