CH2=CHCH2CN Database
货号: | MHC001 |
生长状态: | 存活率95% |
年限: | 6个月 |
运输方式: | 常温运输 |
器官来源: | 肝Liver |
是否是肿瘤细胞: | 否 |
细胞形态: | 梭形,贴壁 |
免疫类型: | NC |
物种来源: | 小鼠Mouse |
相关疾病: | NC |
组织来源: | 肝脏Liver |
品系: | NC |
ATCCNumber: | NC |
细胞类型: | Adherent |
肿瘤类型: | NC |
供应商: | biocyto |
数量: | 10^6 |
规格: | 10^6 |
Primaryhepatocytecultureisavaluabletoolthathasbeenbroadlyusedinbasicresearchofliverfunction,disease,pathophysiology,pharmacologyandotherrelatedsubjects.Themethodbasedontwo-stepCollagenaseperfusionforisolationofintacthepatocyteswasfirstintroducedbyBerryandFriendin1969and,sincethen,hasundergonemanymodifications.ThemostcommonlyusedtechniquewasdescribedbySeglenin1976.
Product | Catalogno. | Amount | Storage |
MousePrimaryHepatocytes | RHC001 | 1X10^6/vial | inliquidnitrogen |
ProductUse
ForResearchUseOnly.Notforuseindiagnosticprocedures
ImportantInformation
Hepatocytecellgrowbestmediumsupplemnetedwith10%FBSinM199Media
CultureConditions
CultureType:Adherent
TemperatureRange:36℃to38℃
IncubatorAtmosphere:Humidifiedatmosphereof5%CO2
Size:Thetypicalhepatocyteiscubicalwithsidesof20-30µm
PassagingAdherent Cells
Allsolutionsandequipmentthatcomeincontactwiththecellsmustbesterile.
Always usepropersteriletechniqueandworkinalaminarflowhood.
1.Removeanddiscardthespentcellculturemediafromtheculturevessel.
2.Washcellsusingabalancedsaltsolutionwithoutcalciumandmagnesium (approximately2mLper10cm2culturesurfacearea).Gentlyaddwashsolutionto thesideofthevesseloppositetheattachedcelllayertoavoiddisturbingthecell layer,androckthevesselbackandforthseveraltimes.
Note:Thewashstepremovesanytracesofserum,calcium,andmagnesiumthat wouldinhibittheactionofthedissociationreagent.
3.Removeanddiscardthewashsolutionfromtheculturevessel
4.Addthepre-warmeddissociationreagentsuchastrypsinorTrypLE™tothesideof theflask;useenoughreagenttocoverthecelllayer(approximately0.5mLper10 cm2).Gentlyrockthecontainertogetcompletecoverageofthecell layer.
5.Incubatetheculturevesselatroomtemperatureforapproximately2minutes.
Note: thattheactualincubationtimevarieswiththecelllineused.
6.Observethecellsunderthemicroscopefordetachment.Ifcellsarelessthan90% detached,increasetheincubationtimeafewmoreminutes,checkingfordissociation every30seconds.YoumayalsotapthevesseltoexpeditecellTetachment.
7.When≥90%ofthecellshavedetached,tiltthevesselforaminimallengthoftime toallowthecellstodrain.Addtheequivalentof2volumes(twicethevolumeused forthedissociationreagent)ofpre-warmedcompletegrowthmedium.Dispersethe mediumbypipettingoverthecelllayersurfaceseveraltimes.
8.Transferthecellstoa15-mLconicaltubeandcentrifugethenat200×gfor5to10 minutes.Notethatthecentrifugespeedandtimevarybasedonthecelltype.
9.Resuspendthecellpelletinaminimalvolumeofpre-warmedcompletegrowth mediumandremoveasampleforcounting.
10.Determinethetotalnumberofcellsandpercentviabilityusingahemacytometer, cellcounterandTrypanBlueexclusion,ortheCountess®AutomatedCellCounter.If necessary,addgrowthmediatothecellstoachievethedesiredcellconcentrationand recountthecells.
Fuction
Ahepatocyteisacellofthemainparenchymaltissueoftheliver.Hepatocytesmakeup70-85%oftheliversmass.Thesecellsareinvolvedin:
Proteinsynthesis
Proteinstorage
Transformationofcarbohydrates
Synthesisofcholesterol,bilesaltsandphospholipids
Detoxification,modification,andexcretionofexogenousandendogenoussubstances
Initiationofformationandsecretionofbile
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发布于 : 2018-06-09
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