ProductDescription
Extralink®Lite(PEGDA,polyethyleneglycoldiacrylate)0.5mLvialscontain2.5mgofdiacrylatedPEGand2.5mLvialscontain12.5mg.Vialsareblanketedbynitrogenandunderaslightvacuum.
Extralink®ispolyethylenediacrylate(PEGDA)withamolecularweightof3500Da.Itisathiol-reactivecrosslinkerthatconvalentlyreactswiththethiolgroupsinGlycosil®,Heprasil®andGelin-S®toformviscoelastichydrogels.Extralink®Litevialsare>80%acrylatedandsuppliedasthecrosslinkingcomponentintheHyStem®hydrogelkits.GelationExtralink®LiteisusedtochemicallycrosslinkGlycosil.Thegelationtimerangesfromasshortas5minutestoaslongasacoupleofhours,dependingupontheamountofExtralinkusedandtheconcentration/dilutionoftheGlycosil.Extralink®LiteisusedinplaceofExtralinkforapplicationswhereGelin-Sisnotincluded.ThehigherrelativeconcentrationofGlycosilrequireslesscrosslinkertoemulatethegelationtimeandstiffnessofHyStem-CandHyStem-HP.
STORAGE
Extralink:StoreExtralinkintheoriginalvialunopenedat-20°Cforuptooneyear.ReconstitutedExtralinksolutionscanbestoredat-20°Cfor~onemonth.
Note:Itisrecommendedtoreconstituteeachvialinitsentirety.
DirectionsforUse
Whenreconstituted,itwillbein1xphosphatebufferedsaline(PBS)bufferpH~7.4.TheamountofDGWaterusedfordissolutiondependsonthevial.
Extralink®shouldbepreparedinthefollowingmanner:
- AllowtheExtralink®vialtocometoroomtemperature.
- Underasepticconditions,usingasyringeandneedle,addtothevialtheamountofDGWaterindicatedonthelabel.
- Invertseveraltimestodissolve.
- Extralink®isusedtochemicallycrosslinkhydrogelsmadefromGlycosil®orHeprasil®andGelin-S®.Extralinkdoesnotformahydrogelonitsown.
- Typically,Extralink®isusedina4:1volumeratiowithGlycosil®orHeprasil®asfollows:
- 0.25mLExtralink®iscrosslinkedwith1mLGlycosil®
- 0.25mLExtralink®iscrosslinkedwith1mLHeprasil®
- NotethegelationtimevariesdependingupontheamountofExtralink®,theamountofGlycosil®orHeprasil®used.
Note:Gelin-SwillnotformahydrogelwhenmixedwithExtralink®withoutGlycosil®orHeprasil®.
Note:HydrogelsmadeusingonlyExtralink®andGlycosil®orHeprasil®willnotsupportcellattachment.Forcellattachment,theGelin-Scomponentorotherbindingsitesisrequired.
ProductQ&A
Globularparticleslessthan75kDashouldbeabletofreelydiffusethroughaHyStemhydrogel.
WhenreconstitutedusingDGwater,thepHofeachHyStemcomponentwillbeapproximately7.4-7.6.
Oneyearfromthedateofreceipt,ifstoredproperly.
Anysterile,deionized,degassedwatercanbesubstitutedforreconstitution.However,inordertoensureaccurateandpredictabledissolutionandgelationtimes,ourDGWaterishighlyrecommended,asitisdegassed,blanketedinargon,andhasundergonevalidationtestingwitheachHyStemcomponent.
Gelin-Sprovidescellularattachmentsiteswhenincorporatedinthehydrogel.Gelin-Sisthiol-modified,denaturedcollagenI,derivedfromeitherbovineorporcinesources.Gelin-SisincludedinallHyStem-CandHyStem-HPkits.
Gelin-Shasbeenthiol-modifiedinthesamemannerasthehyaluronaninGlycosil(orHeprasil),sothatitcovalentlycrosslinkswiththeExtralinkintheHyStemhydrogels.
Yes.Peptidesthatcontainacysteineresiduecanbeused.Thecysteineresiduemustbepresentforthepeptidetobecovalentlybondedtothehydrogelsubstrate.
Yes.ECMproteins,suchaslaminin,collagen,fibronectin,orvitronectincanbenon-covalentlyincorporatedintothehydrogelpriortocrosslinking.
HyStemhydrogelsandspongesdifferinhydrationandhomogeneity.HyStemspongesaretypicallypolymerizedhydrogelsthataresubsequentlyfreeze-dried.Theresultingspongeisafibrous,meshnetworkwithporesandnichesthatenablecellstoinfiltrateandadhere.AtrueHyStemhydrogelisanencapsulatingliquidthatpolymerizesaroundsUSPendedcellsinculture.
No.ThecomplianceofthehydrogelsissetbytheamountofExtralinkcrosslinkeradded,theconcentrationofGlycosil(orHeprasil)andGelin-Sused,andtheratioofGlycosil(orHeprasil)toGelin-S.Oncethischemicalstructureofthehydrogelisfixed,itisnotalteredbyprolongedexposuretocellculturemedium.
HyStemspongescanbeterminallysterilizedbyE-beam.HyStemhydrogelshavenotyetbeenvalidatedforusewithE-beamsterilizationmethods.HyStemhydrogelsarenotterminallysterilizedbygammairrADIation.
Gelationtimeisaffectedbymultipleaspectsofthegel’scomposition.Onewaytochangethegelationtimeofahydrogelistovarytheamountofcrosslinkerused.GelswithaloweramountofExtralinkcrosslinkerwillhavealongergelationtimethanthosewithahigheramountofcrosslinker.Changingtheamountofcrosslinkerwillproduceslightchangesingelationtime.GelationtimecanbedramaticallychangedbyvaryingtheGlycosil(orHeprasil)andGelin-Sconcentrations.ConcentratedsolutionsofGlycosil(orHeprasil)andGelin-Swillcreateasolutionwithamuchshortergelationtime.ThiscaneasilybedonebyreconstitutingthecomponentsinasmallervolumeofDGWater.Alternatively,dilutingthesecomponentsinlargervolumesofDGWaterwilldramaticallyincreasethetotaltimetoformthehydrogel.
HyStemHydrogelsarevirtuallytransparentandshouldnotinterferewithmicroscopy.
HyStemhydrogelsmaygeneratemildinflammationaspartofthebody’snaturalhealingprocessinresponsetoinjury.HyStemhydrogelsdonottriggerimmuneresponsewhenusedinvivo.(Theseproductsarenotforhumanuse)
HyStemisdegradedinvivobymatrixmetalloproteinases(Collagenases)andhyaluronidases.
Trypsin,Dipase,collagenase,andhyaluronidasehavebeenusedtohelpdetachcellsfromthesurfaceorfromwithinHyStemhydrogels.
Ingeneral,theporesizeforHyStem-CandHyStem-HPhydrogelsis~17nm.
ProductApplications
Clickonthetitleofthedesiredprotocoltolearnmore:
2DCellGrowthonHyStemHydrogels
HyStem3DCellEncapsulationforCellDeliveryApplicationsGuide
HyStem3DCellEncapsulationinhydrogelsusing96-wellplates
HyStem3DCellEncapsulationinhydrogelsusingTCInserts
EnzymeDigestionofHyStemHydrogelsforRecoveryofEncapsulatedCells
FluorescentLabelingofHyStemHydrogels
CellRecoveryfromSurfaceofHyStemHydrogels
HyStemECMIncorporation
HyStemGelationTimeVariation
HyStemStiffnessVariationProtocolfor7.5mLkit
HyStemStiffnessVariationProtocolfor12.5mLkit
ProductCertificateofAnalysis
SafetyandDocumentation
CertificateofOrigin
SafetyDataSheet
ProductDisclaimer
ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.
