Biosilk

A 3D culture environment for expansion and differentiation of human PSCs. 

Biosilk is a biomaterial made from recombinant spider silk protein for 3D cell culture. The Biosilk solution may advantageously be functionalized with the Biolaminin product of choice, for improved cell functionality. 

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750 µg 1 x 250 µL €240
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A biomaterial designed for 3D cell culture of human pluripotent stem cells 

Biosilk is a biomaterial made from recombinant spider silk protein for 3D cell culture. The Biosilk solution may advantageously be functionalized with the Biolaminin product of choice, for improved cell functionality. Biosilk is ideal for integration, proliferation and subsequent lineage-specific differentiation of human pluripotent stem cells in a 3D format.

Biosilk 521 foam and fibers


Biosilk is a biocompatible, biodegradable and non-immunogenic biomaterial, which facilitates the use in clinical applications. The extendable Biosilk macrofibre material can self-assemble into several formats; foams, fibre bundles and  thin films. 

 

A MORE BIOLOGICALLY RELEVANT CULTURE ENVIRONMENT 

Compared to 2D cultures, a 3D culture is a more biologically relevant culture environment with a higher degree of architectural complexity that retains homeostasis for longer.

 

Biosilk 521 is a biorelevant cell culture system


Contrary to many other 3D scaffolds, the cells can be added to the Biosilk solution before the network stabilizes, resulting in a uniform integration of the cells between the microfibers. Unlike cells cultured in hydrogel, cells seeded in the Biosilk scaffold attach along the microfibers and are in contact with other cells like in tissue. Moreover, the cells proliferate more efficiently and migrate in the Biosilk scaffold and have full and even access to both the medium, matrix and neighbouring cells, compared to when encapsulated in a hydrogel.

FEATURES AND SPECIFICATIONS:

  • Flexible 30 culture system that can be assembled into different formats

  • Elastic microfibers mimicking the ECM

  • Biocompatible

  • Biodegradable

  • Non-immunogenic

  • Defined and animal component-free to the primary level

  • For research use only


 

 

 

BIOSILK 521 FOAM MORPHOLOGY

Representative pictures of the Biosilk 521 foam taken from above day 0, 1 and 3 after cell seeding. The initial small bubbles merge into bigger bubbles that disperse after a few days, resulting in a 3D scaffold with integrated cells. 

 Biosilk 521foam morphology

 

PROTOCOL

Preparation of Biosilk solution

  1. Thaw the Biosilk solution at RT without moving the vial.
  2. Add Rock inhibitor to the thawed solution to final concentration of 10 µM. Gently pipette 3 times.

Note:
!   Do not vortex or shake the vial and be careful when mixing to avoid introduction of air bubbles.
!   It will take around 10 min for the frozen Biosilk solution to thaw at ambient temperature. The thawed solution should be used as soon as possible, within1 hour at the latest. The thawed Biosilk will gradually turn milky in ambient room temperature.



Preparation of a concentrated cell suspension

  1. Prepare a concentrated single cell suspension according to “Instruction For Use BL003”, roughly 10.000-15.000 cells/µL, diluted in warm media supplemented with 10 µM ROCK inhibitor.

Note:
!   The cell suspension should be prepared freshly for foam seeding (use within 20 min after detachment from plate). Cell suspension standing too long in RT will result in reduced cell amplification rate in the foam.

 

Integration and expansion of hPSC in Biosilk foam

  1. Transfer 20 µL of the prepared Biosilk solution from step 1 to the center of one culture well.
  2. Use a pipette with a tip for 200 µL and set at 40 µL. Push air bubbles into the droplet by quickly pipetting up and down 20 strokes, thereby creating a dense foam. Spread out the foam in circular motions with the pipette tip during pipetting to an area covering 0.7-1 cm in diameter.
  3. Immediately add 1–5 µL (typically 30.000-60.000 hPSCs/foam) of the cell suspension from step 2 (volume ratio of cell to Biosilk ≤0.25). Use the pipette set at 40 µL with a new tip and disperse the cells throughout the 3D structure by 5 additional strokes.
  4. Repeat step 1 to 3 to create the desired number of foams. One vial of Biosilk (270 µL) is sufficient for 12 foams.
  5. Place the plate with the cell-containing foams in an incubator at 37°C for 20 min. During this time, the 3D structure is stabilized. 
  6. Gently add 0.7-1 mL of the pre-warmed medium containing 10 µM ROCK inhibitor per well, enough to cover the foam.
  7. Place the plate back into the incubator.
  8. Feed the cells daily with fresh culture media without ROCK inhibitor.

Note
!    For best foam stability, cell suspension for foam seeding could be prepared beforehand or during the Biosilk thawing time.
!    It takes approximately 1 min to generate each cell-containing foam. It’s good to plan the timing for best cell and foam quality.

4. Differentiation

Differentiation protocol of interest can be used when desired confluency has been reached.

IMPORTANT NOTES

  • All steps must be carried out under aseptic conditions

  • Biosilk should be stored at -80°C

  • Thaw the Biosilk solution at RT without moving the vial

  • Do not vortex or shake the vial and be careful when mixing to avoid introduction of air bubbles

  • Thawed Biosilk solution has to be used within 1 hour. Re-freezing or storage in fridge is not recommended and will result in decreased foaming efficacy

  • For research use only

Product name

Biosilk

 

Product code

BS-0101

 

Declaration

For research use only

 

Product description

Recombinant spider silk protein for 3D culture applications

 


Documents

MSDS

 

Storage

-80°C

 

Stock concentration

0.1mg/ml

 

Stability

12 months 

 

Shipping condition

Dry Ice

 

Appearance

Clear, colourless

 

Product application

Human PSC expansion and differentiation  

 

Classification

Defined and animal component-free, human recombinant protein