3D Scaffolds

Next Generation Jellyfish Collagen 3D Scaffolds for in vitro cell culture and tissue engineering.

Jellagen® 3D Scaffolds offer the potential for cells to possess a natural tissue-like structure offering better function in a more physiologically relevant manner.

Jellagen® 3D Scaffolds create a 3D cell culture environment that enables cells to maintain their in vivo morphology, behaviour and responsiveness within an in vitro model system. The porous network within the sponge allows for efficient nutrient uptake and increased surface area for cell attachment and migration.

The grade of Jellagen® jellyfish collagen used to manufacture these scaffolds has been tested to verify its applicability for routine cell culture research using human primary and iPSC-derived cell lines. Jellagen® Jellyfish collagen has been shown to promote cellular attachment, proliferation and differentiation to develop functional matrices.

Cell lines that have been cultured successfully on Jellagen® jellyfish collagen include, but are not limited to: Chondrogenic progenitor cells, ovarian cancer cells and iPSC-derived microglia

Features  Benefits
Innovative Offers a viable alternative to mammalian and synthetic scaffolds
Non-mammalian Highly purified jellyfish collagen alternative providing consistent, repeatable results
Batch to batch consistency Offers improved research productivity allowing security of product consistency and reproducible results
Cross-linked EDC cross-linked scaffolds for enhanced mechanical and thermal stability, resorbable in vivo.
Evolutionary ancient collagen demonstrating sequence homology to collagen I, II & V Universal applications for multiple cell types and regenerative medicine
Manufactured to ISO13485 Follows a quality controlled manufacturing producing a consistent scaffold
Uniform pore size Promotes cell seeding, invasion, proliferation and differentiation. Allows for growth factor permeation and gas exchange, ensuring long term cell survival.
Natural scaffold Jellagen® 3D Scaffolds exhibit similar physiological components and properties to the ECM of the in vivo micro environment

Product numbers 

  • JSM96F, JSM96H, JSM96Q
  • JSM48
  • JSM24F, JSM24H
  • JSM06
    *Bespoke and non cross linked formats available on request and subject to volume
    (F) Full, (H) Half, (Q) Quarter

Jellagen® 3D Scaffolds

 

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PRODUCT INFORMATION JELLAGEN® - 3D SCAFFOLDS
Format 96, 48, 24, & 6 - well scaffolds, cast in plates
Collagen Jellyfish collagen
Storage Room temperature
Serum level Serum free
Shelf life Under evaluation
Plate polymer Non-tissue culture treated, polystyrene and non-pyrogenic
Colour White to off-white
Bioburden Negative
Shipping conditions Room temperature
pH Neutral

 

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How to use Jellagen 3D Scaffolds

Characterisation 

Dimensions:
• 96 well: 5mm x >0.5mm
• 48 well: Under development
• 24 well: Under development
• 6 well: Under development
Bespoke sizing also offered

pH: Approximately 7.0 to 7.4 when suspended in PBS or tissue culture media
Storage/Stability: Room Temperature – Heating above 40ºC is not recommended.
Store in a cool, dry place. The stability of the product is under evaluation.

Precaution and 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. The procedure below is provided as a guideline, but the onus is on the end-user to tailor the conditions of their experiment to their needs and that of their cell line.

Preparation and seeding 

Note: Cell attachment to the scaffold is generally the most critical step in tissue culture. Temperature, pH, gas exchange and cell concentration can affect the rate and efficiency of attachment. Optimum seeding rate depends on the type of cell being cultured.

Steps:

1. Using aseptic technique, remove the scaffold plate from the packaging in a laminar flow environment.

2. Jellagen scaffolds are packaged in a non-tissue culture plate and as such can be used immediately. However, should the scaffold need to be moved, do so with a sterile instrument and take care not to damage the scaffold as it is being transferred.
Note: Tissue-culture treated plasticware may need to be coated with 2% agarose to prevent cell attachment to the plastic instead of the scaffold.

3. Suspend cells at desired concentration in media and dispense sufficient volume of cell solution on top of the scaffold placed in the well.

4. Transfer to a 37ºC incubator for about 1 – 2 hours to allow for initial cell attachment.

5. After 24 hours, remove the plate from the incubator and check for cell attachment. Additional testing may be required to optimise the time it takes for the cells to attach to the scaffold. Check the morphology of the cells. Cell adherence and spreading will dictate the time for attachment.

6. Once the cells have adequately attached to the scaffold, increase the final volume in each well to fully cover and provide adequate medium for the culture system.

Changing the media 

Change the media 12 to 24 hours after the initial seeding. The frequency of changes will be determined by cell type, cell attachment efficiency, pH utilisation of medium nutrients available to cultures. More frequent medium changes may be required compared to 2D culture systems.

Harvesting of cells 

Note: Digestion with proteases such as trypsin, papain (cysteine protease)1 or collagenase are suitable methods of releasing cells from the Jellagen-3D Scaffolds. The strength of the attachment of the cells to the collagen scaffolds will vary from cell line to cell line. The enzyme concentration and digestion time will vary depending upon the activity of the enzyme and the confluency of the cells. Collagenase and/or trypsin may be the preferred method. If using papain, the following method is suggested:

1. Prepare enzyme buffer solution (20mM NaAc pH 6.8, 1mM EDTA, 2mM DTT, 330μg/ml papain)

2. Washing the scaffold with EDTA-PBS may assist the protease digestion. Add sufficient volume to cover the scaffold.

3. Aspirate the EDTA-PBS solution from the well.

4. Add sufficient dissociation solution to the well to fully over the scaffold.

5. Transfer to a 37ºC incubator. Check for cell detachment periodically for cell detachment.

6. Once the cells have fully detached, remove the cells and dispense in a centrifuge tube.

7. Centrifuge the cells as required.

Useful references 

1. Estes, B. T. & Guilak, G. 2011. Three-dimensional culture systems to induce chondrogenesis of adipose-derived stem cells. Methods in Molecular Biology, 702.
2. Eun Song., So Yeon Kim., Taehoon Chun., Hyun-Jung Byun. & Young Moo Lee. 2006. Collagen scaffolds derived from a marine source and their biocompatibility. Biomaterials 2. 2951–2961
3. Sewing, J., Klinger, M. & Notbohm, H. 2015. Jellyfish collagen matrices conserve the chondrogenic phenotype in two- and three-dimensional collagen matrices. Journal of Tissue Engineering and Regenerative Medicine.
4. Carletti, E., Motta, A. & Migliaresi, C. 2011. Scaffolds for Tissue Engineering and 3D Cell Culture. Methods in Molecular Biology. Humana Press.
5. Chan, B. P. & Long, K. W. 2008. Scaffolding in tissue engineering: general approaches and tissue-specific considerations. European Spine Journal, 17, 467-479.
6. Hoyer, B., Bernhardt, A., Lode, A., Heinemann, S., Sewing, J., Klinger, M., Notbohm, H. & Gelinsky, M. 2014. Acta Biomaterials. Feb;10(2):883-92.
7. Fuss, M., Ehlers, E. M., Russlies, M., Rohwedel, J. & Behrens, P. 2000. Characteristics of human chondrocytes, osteoblasts and fibroblasts seeded onto a type I/III collagen scaffold under different culture conditions. A light, scanning and transmission electron microscopy study. Annals of Anatomy, 182, 303-310.
8. Bernhardt, A., Paul, B. & Gelinsky, M. 2018. Biphasic Scaffolds from Marine Collagens for Regeneration of Osteochondral Defects. Marine Drugs. 16, 91.

FRM-95 Rev 00

 

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