ProductDescription
ForprintingwithLifeink®200,wehighlyrecommendusingtheFRESHprintingtechniqueandprintingat2-8°C.Threedimensional(3D)gelsallowforthestudyoftheeffectsofthemechanicalpropertiesoftheextracellularmatrix(ECM),suchasdensityandrigidity,oncelldevelopment,migration,andmorphology.Unlike2Dsystems,3Denvironmentsallowcellextensionstosimultaneouslyinteractwithintegrinsonallcellsurfaces,resultingintheactivationofspecificsignalingpathways.Gelstiffnessorrigidityalsoaffectscellmigrationdifferentlyin3Dversus2Denvironments.
Abioinkthatresembleslifeshouldincludethefollowingtraits:HighPrintResolution,shear-thinning,strongmechanicalstrength,superiorcytocompatibility,allowforcellularremodeling,andisbiomimetic.Lifeink®200fulfillsthoserequirements.
AdvancedBiomatrixoffersLifeink®200,abioinkthatisa35mg/mltypeIcollagen.Lifeink®200ispHneutralcollagenwithphysiologicalsaltconcentration.Thecollagenstartingmaterialispurifiedusingacontrolledmanufacturingprocess.
ReadthegroundbreakingpublicationusingLifeink®200forbioprintingheartcomponentshere.
Lifeink®200bioinkhasthefollowingcharacteristicsasshownbelow:
| Parameter,Testing,andMethod | Lifeink®200#5278 |
| SterilizationMethod | Filtration,AsepticProcessing |
| Sterility-USPmodified | Nogrowth |
| Form | OpaqueGel/Paste |
| PackageSize | 5mL |
| StorageTemperature | 2-8°C |
| PrintTemperature | 2-8°C |
| ShelfLife | Minimumof6monthsfromdateofreceipt |
| Endotoxin-LAL | <10.0EU/mL |
| pH | 6.9-7.6 |
Osmolality(mOsmoH2O/kg) | 290-370 |
SourceMaterial | BovineCollagen |
ElectrophoreticPattern-CoomassieBlue | Characteristic |
CollagenConcentration-Biuret | 30-45mg/mL |
CollagenPurity-SilverStaining | >99% |
ContinuousFlowExtrusion | Pass |
DirectionsforUse
DownloadthefullPDFversionorcontinuereADIngbelow:
Note:Employasepticpracticestomaintainthesterilityoftheproductthroughoutthepreparationandhandlingofthecollagenandothersolutions.
Note:Itisrecommendedthatthecollagenandotherworkingsolutionsbechilledduringthepreparationofthecollagen.
Note:Vortexingofcollagenisnotrecommendedatanystep.
Note:EnsurethatNObubblesenterthesystem.Bubblesinthesystemwhilemixingwillturnyourinkintoafoam-likematerial.
Note:WerecommendprintingLifeink®at2-8°Cintoagelatinslurrysupport(FRESHmethod)forbestresults.
CellAdditiontoBioink:Note:Keepcollagenchilledthroughouttheseprocesses
- Addtheconcentratedcellsuspensionincellculturemediatoasterilesyringe.
Videoforstep1
Note:Werecommendafinalbioinkcellconcentrationof5X106/mLorgreater.
Note:Forbestresults,add2mLofcellsuspensionper5mLofcollagenBioink.Useasimilarratioforsmallervolumes.
- Placesterilecouplerontheendofthesyringewiththecellsuspension.
- Slowlypushplungerinuntilmediaformsaslightexternalmeniscusabovetheendofthecoupleronthesyringe.
- Removecapfromthesyringewithcollagenandslowlypushplungerinuntilcollagenformsaslightexternalmeniscusabovetheendofthesyringe.
- Couplethesyringewithcellstothesyringewithcollagen.(Ensurethattherearenoairbubblesinthesystem.The“externalmeniscus”onbothsyringeshelpsensurethattherearenoairbubblesintroduced).
- Slowlypushplungersbackandforth~40timestoensurethoroughmixing.Endwithallofthematerialinthesyringetobeusedforprinting.
Videoforsteps2-6
- Thecell-ladenbioinkisnowreadyforpositivedisplacementprinters.Allow5-10minutesforthesyringetoequilibratebackto2-8°Cbeforeprintingforbestresults.
Note:Forpneumaticprinters,transferthecollagenintoanappropriatesyringeusingthecoupler.Thenewsyringeshouldhavethesealinserted,buttheplungerremoved.Centrifugethesyringeat2000RPMfor1minuteaftertransferringthecollagentoremoveanyairbubbles.Donotdothecentrifugestepiftherearecellsalreadymixedin.
Watchthisvideoforhelp
GeneralPrintingNotes:
- Touseasmallervolumeofcollagen,simplytransferthedesiredamountofcollagentoanothersyringe,usingtheprovidedsterilecoupler.Toremovetheairfromthenewsyringe,youcandoeitherofthefollowing:
- Centrifugethesyringe(capped)withthecappointinguptocausetheairtoaccumulateatthecap.Evacuatetheair.
- Centrifugethesyringe(capped)withthecappointingdown,andthenuseahemostattosqueezethesyringewhilepushingtheplungertoallowtheairtoescape.
Removingairwithahemostatvideo
- Completeyourprintwithin1hourofcelladditiontosyringe/bioinkforbestcellviABIlityresults.
- WhenprintingwithFRESHgelatinslurry,allowthefinalprintedstructuretoincubateat37°Cfor30to60minutesandthenreplacethegelatinwithmedia.
- Avoidbubbles.
ProductCellAssay
FibroblastsDay2
AdiposeDerivedStemCellsDay1
AdiposeDerivedStemCellsDay6
Fibroblastsday1
Fibroblastsday5
ProductBioprints
FullScaleHumanNeonatalHeart
HeartValve-Closed
HeartValve-Open
Meniscus
SimpleScaffold
Justahand...
(Canyouseethesinglelayerthinfingernails?)
ProductReferences
ReferencesforLifeink®collagenbioinks:
Lee,A.etal.3Dbioprintingofcollagentorebuildcomponentsofthehumanheart.Science365,482–487(2019).
Maxson,EvaL.,etal."Invivoremodelingofa3D-Bioprintedtissueengineeredheartvalvescaffold."Bioprinting(2019):e00059.
Filardo,G.etal.Patient-specificmeniscusprototypebasedon3Dbioprintingofhumancell-ladenscaffold.Bone&JointResearch8,101–106(2019).
Schmitt,T.AnalysisandClassificationof3-DPrintedCollagen-BioglassMatricesforCellularGrowthUtilizingArtificialNeuralNetworks.UniversityThesis(2018).
Balakhovsky,Y.M.,Ostrovskiy,A.Y.&Khesuani,Y.D.EmergingBusinessModelsTowardCommercializationofBioprintingTechnology.3DPrintingandBiofabrication1–22(2017).doi:10.1007/978-3-319-40498-1_25-1
Fox,S.etal.Asimplifiedfabricationtechniqueforcellularizedhigh-collagendermalequivalents.BiomedicalMaterials14,041001(2019).
ProductCertificateofAnalysis
SafetyandDocumentation
SafetyDataSheet
CertificateofOrigin
DeclarationofMaterialSource
ProductDisclaimer
ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.
