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TransfectionReagentforAstrocytes(PrimaryAstrocyte Cells,Astroglia)
TransfectionEfficiency:
Reagentexhibitsatleast70%transfectionefficiencyofsiRNAdelivery.TransfectionefficiencywasdeterminedbyqRT-PCR.
TransfectionProtocolandMSDS:
DownloadAltogenBiosystemsAstrocyteTransfectionProtocol:[PDF]
DownloadMSDS:[PDF]
AstrocyteCells(PrimaryAstrocytesandAstroglia):
Thehumanbraincomprisestwomajorcelltypes:neuronsandglia.NeuronsareresponsIBLefortransmittingnervesignalswhilegliasafeguardneurons.Astrocytes,asortofglialcells,makeover50%ofallbraincells.IntheeventofanydistresstotheCentralNervousSystem(CNS),aprocesscalledgliosisistriggered.Gliosisinvolvestherapiddifferentiationofglialcellstoprovidemechanicalsupporttoneurons.However,excessivegliosisissymptomaticofvariousformsofbraincancer.PerthestudypublishedintheJanuary2016issueofTumorBIOLOGy,astrocytesplayanimportantroleincontrollingtheenvironmentaroundatumor.Astrocytesretardthedetectionofcancercellsbytheimmunesystemandhelpenhancetheirproliferation.Preclinicalresearchtargetingastrocytescanprovidevaluableinsightintomolecularpathwaysthatcancurbtumors.Transfectionisawell-regardedtechniquetoconductpreclinicalresearchinmanydifferentcells,includingastrocytes.
Astrocytesarestar-shapedglialcellsinthebrainandspinalcord.Astrocytesaresupportivecells,andtheirfunctionsincludebiochemicalsupportofendothelialcells,whichformtheblood-brainbarrier,providingnutrientstothenervetissue,extracellularK+levelregulation,neurotransmittersremoval,andhelpingrepairthebrainandspinalcordfollowingtraumaticinjuries.Theyarealsodependentonmyelinationandsynapsesformation.ManyprimaryastrocytesexpresstheintermediatefilamentglialfibrillaryacidicproteinorGFAP.Researchindicatesthatastrocytescommunicatewithneuronsthroughthereleaseofgliotransmittersviaacalciumionprocess.Therearethreetypesofastrocytesinthecentralnervoussystem:fibrous,protoplasmicandrADIal.Itisbelievedastrocytescellbodiesareseparateanddonotoverlap.
Astrocytes,alsoknownasastroglia,arecellsinlivingorganismsthatmakeuppartsofthenervoussystem,particularlyinthebrainandspinalcord.Theyworkwiththebody’snaturalresponsetotraumainthebrainandspinalcord,byrepairingandprovidingnutrientstoaffectedareas.Becauseofthis,astrocytesmaybeusefulinthestudyofspecificmedicaltreatmentsforpatientssufferingfromspinalcordinjuries.Thesecellsarecrucialtoresearchinthefieldofneuroscience.
Primarycellculturesareusedinbiologicalandgenetherapystudiesandserveasvaluablemodelsystemsthatmaymoreaccuratelyrepresentthebiologyofhealthycells.Manyculturedcelllines,aswellasthemajorityofprimarycellcultures,canbetransfectedwithexogenousnucleicacidswhenappropriatetransfectionapproachesareemployed.Sincethemajorityoftransfectionmethodscausessignificanttoxicityinprimarycellcultures,optimizingthisprocedure,specificallytheprotocolandreagentstobeutilized,isessentialfordevelopinganeffectivetransfectionstrategiesforagivencelltype.ALTOGEN®Kitsforprimarycellsandsensitivecelllineshavebeendesignedtohavesignificantlylowercytotoxicitythanotheralternatives.
Data:
Figure1.SiRNAstargetingLaminA/CmRNAornon-silencingcontrolsiRNAweretransfectedintoactrocytecellsfollowingtherecommendedprotocol.At48hourspost-transfectionthecellswereanalyzedbyqRT-PCRforgeneexpressionlevels.18SrRNAlevelswereusedtonormalizetheLaminA/Cdata.Valuesarenormalizedtountreatedsample.Dataaremeans±SD(n=3).
Figure2.ProteinexpressionofLaminA/CinAstrocytecells.DNAplasmidexpressingLaminA/CorsiRNAtargetingLaminA/CweretransfectedintoAstrocytecellsfollowingAltogenBiosystemstransfectionprotocol.At72hourspost-transfectionthecellswereanalyzedbyWesternBlotforproteinexpressionlevels(normalizedbytotalprotein,10µgoftotalproteinloadedpereachwell).Untreatedcellsusedasanegativecontrol.
SelectedAstrocyteTransfectionReagent productcitations:
AltogenBiosystems:
AltogenBiosystemsprovidespre-optimizedtransfectionkitsandelectroporationproductsforlifesciencesandcancerresearch.Transfectionprotocolsareoptimizedforindividualcancercelllines.AltogenBiosystemsdevelopedtwotypesofinvivodeliverykits(animaltransfection):1)Tissue-targetedreagents(deliveryofproteins,DNA,andRNAintoliver,pancreas,orkidneytissues),and2)Broadrangeinvivobiodistributionreagents(PEG-Liposomebasedreagent,Nanoparticle-basedinvivoreagent,Lipid-basedtransfectionkit,andPolymer-basedkit).Advancedformulationofreagentsandoptimizedtransfectionprotocolsprovideefficientintracellulardeliveryofproteins,DNA,mRNA,shRNA,siRNA,andothernegativelychargedbiomoleculesinvitroandinvivo.Readmore abouttransfectiontechnologyatAltogen’sTransfectionResource.
AltogenResearchServices:
AltogenLabsprovidesGLP-compliantcontractresearchstudiesforpre-clinicalresearch,INDapplications,anddrugdevelopment.BiologyCROservices include:Xenograftmodels(30+),developmentofstablecelllines,ELISAassaydevelopment,cell-basedandtissuetargetedRNAistudies,safetypharm/toxassays,andotherstudies(visitAltogenLabs.com).
VolumeOptions:
SouthernBiotech/Goat Anti-Human Lambda-Alexa Fluor® 555/2070-32/1.0 mg
vectorlabs/Biotinylated Aleuria Aurantia Lectin (AAL)/B-1395/1 mg
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Mouse IgG)/PK-6102/1 kit
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Universal)/PK-6200/1 kit
vectorlabs/Unconjugated Musa Paradisiaca (Banana) Lectin (BanLec)/L-1410/5 mg
GiottoBiotech/Calmodulin N60D/5 mg/G02CLM60cn