ProductDescription
ElectrospunGelatinDiscsarefabricatedusingelectrospinningtechniquestocreateaconstructwithauniquestructuresuitableasabiomimeticthree-dimensional(3D)scaffold.Theproductcontainsoverlaidelectrospunfilamentscreatingaporousnetwork,whichpermitscellsandnutrientstoflowcompletelythroughtheporesandprovidesanincreasedsurfaceareaforcellattachment,growthandmigration.
ElectrospunGelatinDiscsarelightlycross-linkedforincreasedmechanicalstrengthanddurABIlityforshortandlong-termtissuecultureexperiments,yetisstillbiodegradableoverthelonger-term.Theaveragefilamentdiametertypicallyrangesfrom500nmto1.5microns.Theelectrospungelatindiscsareapproximately8mmindiameterand1-2mmthick.Theelectrospundiscsfitintoa48wellcultureplateorlarger.Eachpackagecontainsfivediscs.Thisproductissterilizedandready-to-use.
Table1:
Parameter,Testing,andMethod | ElectrospunGelatinDiscs#5214 |
DiscDiameter | 8mm |
PackageSize | 5Discs/Package |
DiscThickness | 0.1-0.2mm |
FiberDiameter | 500nm-1.5um |
StorageTemperature | RoomTemperature |
ShelfLife | Minimumof6monthsfromdateofreceipt |
SterilizationMethod | IrrADIation |
GelatinSource | PorcineGelatin |
DirectionsforUse
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- PreparationandSeeding:
Note:Cellattachmenttothediscisgenerallythemostcriticalstepintissueculture.Temperature,pH,gasexchangeandcellconcentrationcanaffecttherateandefficiencyofattachment.Optimumseedingratedependsonthetypeofcellbeingcultured.
- AsepticallyremovetheElectrospunGelatinDiscsfromthepackaginginalaminarflowworkstation.
- Carefullyplacethediscsintothewellsofa48welltissuecultureplateorlargerusingasterileinstrument.Becarefulnottodamagetheproductasitisbeingtransferred.Itisrecommendedtousenon-treatedtissuecultureplasticware.Note:Tissue-coatedplasticwaremayneedtobecoatedwithagarosetopreventcellattachmenttotheplasticandpromoteattachmenttothedisc.
- SUSPendcellsatdesiredconcentration(approximately5,000cells/cm2)anddispensesufficientvolumeofcellsolutionontopofthediscplacedinthewell.
Note:Avoiddispensingthecellsolutiontoorapidlyasthismaycausedamagetothesponge.
- Transfertoa37ºCincubatorforabout1–2hourstoallowforinitialcellattachment.
- After1–2hour,removetheplatefromtheincubatorandcheckforcellattachment.Additionaltestingmayberequiredtooptimizethetimeittakesforthecellstoattachtothediscs.Checkthemorphologyofthecells.Celladherenceandspreadingwilldictatethetimeneededforattachment.
- Oncethecellshaveadequatelyattachedtothedisc,increasethefinalvolumeineachwelltofullycoverandprovideadequatemediumfortheculturesystem.
- ChangingtheMedia:
- Changethemedia24to36hoursaftertheinitialseeding.Thefrequencyofchangeswillbedeterminedbycelltype,cellattachmentefficiency,andpH.Morefrequentmediumchangesmayberequiredcomparedto2Dculturesystems.
- HarvestingofCells:
Note:Proteasedigestionisthestandardmethodofreleasingcellsfromthediscs.Thestrengthoftheattachmentofthecellstothediscswillvaryfromcelllinetocellline.Theenzymeconcentrationanddigestiontimewillvarydependingupontheactivityoftheenzymeandtheconfluenceofthecells.Collagenaseand/ortrypsinmaybethepreferredmethod.
- WashingthediscswithEDTA-PBSmayassisttheproteasedigestion.Addsufficientvolumetocoverthedisc.
- AspiratetheEDTA-PBSsolutionfromthewell.
- Addsufficientdissociationsolutiontothewelltofullysubmergethediscs.
- Transfertoa37ºCincubator.Checkforcelldetachmentperiodically.
- Oncethecellshavefullydetached,removethecellsanddispenseinacentrifugetube.
- Centrifugethecellsasrequired.
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ProductDisclaimer
ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.