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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