ProductDescription
AdvancedBiomatrixoffersPhotoGel®RUTKit,apurifiedmethacrylatedgelatinkitforvisIBLelight(400-450nm)photocrosslinkablehydrogels.PhotoGel®provides3Dtunablegelswiththeuniqueattributestobepreparedatvariousconcentrationsandcrosslinkedtoprovidevariousgelstiffness.ThisPhotoGel®kitconsistsofpurifiedmethacrylatedporcinegelatinandaVisibleLightPhotoinitiator(400-450nm).ThegelatinistypeA300bloom.
Table1:
Item | CatalogNumber | PackageSize | StorageTemperature |
MethacrylatedGelatin | #5208 | 1gram(2x500mg) | -20°C |
RutheniumPhotoinitiator | #5246 | 100mg | RoomTemperature |
SodiumPersulfatePhotoinitiator | #5247 | 500mg | RoomTemperature |
PhotoGel®isproducedfrommethacrylatedgelatinwherethegelatinhasbeenmodifiedbyreactingthefreeamines,primarilytheε-aminesgroupsofthelysineresiduesaswellasthea-aminesgroupsontheN-termini.>75%ofthetotallysineresiduesofthegelatinmoleculehavebeenmethacrylated.ThephotoinitiatorconsistsofRutheniumandSodiumPersulfatetobeformulatedin1XcellculturemediaorPBS,whichallowsvisiblelightphotocrosslinkingofthecollagenat400-450nm.
Forasepticwork,werecommendfilteringthesolubilizedrutheniumandsodiumpersulfatethrough0.2micronbuttonfilters,asthephotoinitiatorsdonotcomesterile.
Parameter,Testing,andMethod | MethacrylatedGelatin#5208 |
SterilizationMethod | Filtration |
Sterility-USPmodified | Nogrowth |
Form | LyophilizedPowder |
PackageSize | 1gram(2x500mg) |
StorageTemperature | -20°C |
ShelfLife | Minimumof6monthsfromdateofreceipt |
DegreeofMethacrylation | >75% |
GelatinSource | TypeA,300Bloom,Porcine |
HydrogelYoung"sModulusE(Pa) | Characteristic |
DirectionsforUse
DownloadthefullPDFversionorcontinuereADIngbelow:
3DHydrogelPreparation:
Note:Employasepticpracticestomaintainthesterilityoftheproductthroughoutthepreparationandhandlingofthecollagenandothersolutions.
Note:Thefollowinginstructionsarefora10%gelatinmethacrylatesolution.Recommendedconcentrationsare5-20%.
- Warm10mLofwarm,sterile1XPBSor1Xcellculturemediato>60°C.
- Addthe10mL’sofwarmedsolutiontotheambervialcontaining1gramoflyophilizedgelatinmethacrylate.
- Mixonashakertableorrotatorplateuntilfullysolubilized.Keepwarm(>37°C)ifpossible(eg.placeyourrotatorinanincubator)tohelpwithfullsolubilization.
- Calculatethevolumeofphotoinitiatortoaddbymultiplyingthevolumeofsolubilizedgelatinby0.02.Iftheresultingnumberis200ul,forexample,youwilladd200ulofrutheniumand200ulofsodiumpersulfate.
- Solubilizetherequiredamountofruthenium(perstep4)ataconcentrationof37.4mg/mlin1XPBSorcellculturemedia.
- Solubilizetherequiredamountofsodiumpersulfate(perstep4)ataconcentrationof119mg/mlin1XPBSorcellculturemedia.
- Addtherutheniumtothegelatinsolutionandfullymixuntilsolutionishomogeneous.
- Addthesodiumpersulfatetothegelatin/rutheniumsolutionandmixuntilsolutionishomogeneous.
- Addyourcellstothegelatin/photoinitiatorsolution.
- Dispenseyourgelatin/photoinitiator/cellsolutionintothedesireddish(ie.6-wellplate,48-wellplate).
- Forphotocrosslinking,placeprintedstructuredirectlyundera400-450nmvisiblelightcrosslinkingsource.
Longerexposureallowsmorecrosslinking,thougheachcelltypewithstandsdifferentdegreesoflightandfreeradicals(generatedbythephotoinitiator)thatmediatecrosslinking.
Anyexcessmaterialcanberefrigeratedandstored.Thematerialwillgel.Warmbackupto>30°Cforittobecomeliquidagain.Werecommendonlyaddingphotoinitiatortotheamountofgelatintobeusedatthattime.
ProductApplications
PhotoGel®Gelatinmethacrylatecanbeusedtoformcross-linkedhydrogelsfortissueengineering[1]and3Dprinting.Thecommonformsof3DprintingusingLifeink®300includeextrusion[2][3][4],inkjet[5]andphotolithography[6]).
PhotoGel®hasbeenusedforendothelialcellmorphogenesis,[7]cardiomyocytes,[8]epidermaltissue[9],injectabletissueconstructs[10],bonedifferentiation[11],andcartilageregeneration[12].
Gelatinmethacrylatehasbeenexploredindrugdeliveryapplicationsintheformofmicrospheres[13]andhydrogels[14].
References:
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2878615/
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4040163/
- https://www.ncbi.nlm.nih.gov/pubmed/24112804
- https://www.ncbi.nlm.nih.gov/pubmed/20387987
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5380738/
- https://www.ncbi.nlm.nih.gov/pubmed/28349897/
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3643201/
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4551408/
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4608855/
- http://pubs.acs.org/doi/abs/10.1021/bm401533y
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4252258/
- https://www.ncbi.nlm.nih.gov/pubmed/24590160
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4293288/
- http://www.ijpsonline.com/articles/preparation-and-characterization-of-gelatinpolymethacrylic-acid-interpenetrating-polymeric-network-hydrogels-as-a-phsensitive-deli.html
ProductReferences
ReferencesforPhotoGel®:
Rothrauff,BenjaminB.,etal."EfficacyofThermoresponsive,photocrosslinkablehydrogelsderivedfromdecellularizedtendonandcartilageextracellularmatrixforcartilagetissueengineering."Journaloftissueengineeringandregenerativemedicine12.1(2018):e159-e170.
Rothrauff,BenjaminB.,etal."Anatomicalregion-dependentenhancementof3-dimensionalchondrogenicdifferentiationofhumanmesenchymalstemcellsbysolublemeniscusextracellularmatrix."Actabiomaterialia49(2017):140-151.
Rothrauff,BenjaminB.,GuangYang,andRockyS.Tuan."Tissue-specificbioactivityofsolubletendon-derivedandcartilage-derivedextracellularmatricesonadultmesenchymalstemcells."Stemcellresearch&therapy8.1(2017):133.
Bridge,JackChristopher,etal."Electrospungelatin-basedscaffoldsasanovel3Dplatformtostudythefunctionofcontractilesmoothmusclecellsinvitro."BiomedicalPhysics&EngineeringExpress4.4(2018):045039.
Capella-Monsonís,Héctor,etal."Scaffoldsfortendontissueengineering."HandbookofTissueEngineeringScaffolds:VolumeOne.WoodheadPublishing,2019.259-298.
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ProductDisclaimer
ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.