Product Description

Advanced BioMatrix offers PhotoGel® RUT Kit, a purified methacrylated gelatin kit for visible light (400-450 nm) photocrosslinkable hydrogels. PhotoGel® provides 3D tunable gels with the unique attributes to be prepared at various concentrations and crosslinked to provide various gel stiffness.
This PhotoGel® kit consists of purified methacrylated porcine gelatin and a Visible Light Photoinitiator (400-450 nm). The gelatin is type A 300 bloom.

Table 1:

Item Catalog Number Package Size Storage Temperature
Methacrylated Gelatin #5208 1 gram (2 x 500 mg) -20°C
Ruthenium Photoinitiator #5246 100 mg Room Temperature
Sodium Persulfate Photoinitiator #5247 500 mg Room Temperature

 

 PhotoGel® is produced from methacrylated gelatin where the gelatin has been modified by reacting the free amines, primarily the ε-amines groups of the lysine residues as well as the a-amines groups on the N-termini. >75% of the total lysine residues of the gelatin molecule have been methacrylated. The photoinitiator consists of Ruthenium and Sodium Persulfate to be formulated in 1X cell culture media or PBS, which allows visible light photocrosslinking of the collagen at 400-450 nm.

For aseptic work, we recommend filtering the solubilized ruthenium and sodium persulfate through 0.2 micron button filters, as the photoinitiators do not come sterile.

Parameter, Testing, and Method Methacrylated Gelatin #5208
Sterilization Method Filtration
Sterility - USP modified No growth
Form Lyophilized Powder
Package Size 1 gram (2 x 500 mg)
Storage Temperature -20°C
Shelf Life Minimum of 6 months from date of receipt
Degree of Methacrylation > 75%

Gelatin Source

 Type A, 300 Bloom, Porcine

Hydrogel Young's Modulus E (Pa)

 Characteristic

Directions for Use

Download the full PDF version or continue reading below:

3D Hydrogel Preparation:

Note: Employ aseptic practices to maintain the sterility of the product throughout the preparation and handling of the collagen and other solutions.

Note: The following instructions are for a 10% gelatin methacrylate solution. Recommended concentrations are 5-20%.

  1. Warm 10 mL of warm, sterile 1X PBS or 1X cell culture media to >60°C.
  2. Add the 10 mL’s of warmed solution to the amber vial containing 1 gram of lyophilized gelatin methacrylate.
  3. Mix on a shaker table or rotator plate until fully solubilized. Keep warm (>37°C) if possible (eg. place your rotator in an incubator) to help with full solubilization.
  4. Calculate the volume of photoinitiator to add by multiplying the volume of solubilized gelatin by 0.02. If the resulting number is 200 ul, for example, you will add 200 ul of ruthenium and 200 ul of sodium persulfate.
  5. Solubilize the required amount of ruthenium (per step 4) at a concentration of 37.4 mg/ml in 1X PBS or cell culture media.
  6. Solubilize the required amount of sodium persulfate (per step 4) at a concentration of 119 mg/ml in 1X PBS or cell culture media.
  7. Add the ruthenium to the gelatin solution and fully mix until solution is homogeneous.
  8. Add the sodium persulfate to the gelatin/ruthenium solution and mix until solution is homogeneous.
  9. Add your cells to the gelatin/photoinitiator solution.
  10. Dispense your gelatin/photoinitiator/cell solution into the desired dish (ie. 6-well plate, 48-well plate).
  11. For photocrosslinking, place printed structure directly under a 400-450 nm visible light crosslinking source.

Longer exposure allows more crosslinking, though each cell type withstands different degrees of light and free radicals (generated by the photoinitiator) that mediate crosslinking.

Any excess material can be refrigerated and stored. The material will gel. Warm back up to >30°C for it to become liquid again. We recommend only adding photoinitiator to the amount of gelatin to be used at that time.

 

Product Q & A

You can digest it with Proteinase K, though it kills the cells in the process. 

Product Applications

Read our Methacrylated Extracellular Matrices eBrochure Here

PhotoGel® Gelatin methacrylate can be used to form cross-linked hydrogels for tissue engineering[1] and 3D printing. The common forms of 3D printing using Lifeink® 300 include extrusion[2][3][4], inkjet[5] and photolithography[6]).

PhotoGel® has been used for endothelial cell morphogenesis,[7] cardiomyocytes,[8] epidermal tissue[9], injectable tissue constructs[10], bone differentiation[11], and cartilage regeneration[12].

Gelatin methacrylate has been explored in drug delivery applications in the form of microspheres[13] and hydrogels[14].

References:

  1. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2878615/
  2. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4040163/
  3. https://www.ncbi.nlm.nih.gov/pubmed/24112804
  4. https://www.ncbi.nlm.nih.gov/pubmed/20387987
  5. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5380738/
  6. https://www.ncbi.nlm.nih.gov/pubmed/28349897/
  7. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3643201/
  8. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4551408/
  9. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4608855/

Product References

References for PhotoGel®:

 

Rothrauff, Benjamin B., et al. "Efficacy of thermoresponsive, photocrosslinkable hydrogels derived from decellularized tendon and cartilage extracellular matrix for cartilage tissue engineering." Journal of tissue engineering and regenerative medicine 12.1 (2018): e159-e170.

Rothrauff, Benjamin B., et al. "Anatomical region-dependent enhancement of 3-dimensional chondrogenic differentiation of human mesenchymal stem cells by soluble meniscus extracellular matrix." Acta biomaterialia 49 (2017): 140-151.

Rothrauff, Benjamin B., Guang Yang, and Rocky S. Tuan. "Tissue-specific bioactivity of soluble tendon-derived and cartilage-derived extracellular matrices on adult mesenchymal stem cells." Stem cell research & therapy 8.1 (2017): 133.

Bridge, Jack Christopher, et al. "Electrospun gelatin-based scaffolds as a novel 3D platform to study the function of contractile smooth muscle cells in vitro." Biomedical Physics & Engineering Express 4.4 (2018): 045039.

Capella-Monsonís, Héctor, et al. "Scaffolds for tendon tissue engineering." Handbook of Tissue Engineering Scaffolds: Volume One. Woodhead Publishing, 2019. 259-298.

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.