Iwai-Chem/S-Clone SF-O3 (Culture Medium)/Cell Culture/1303

[DevelopmentBackgroundandFeatures]

S-CloneSF-O3isacompletelyserum-freemediumforuseinresearchofmousehematopoieticstemcells.TheproductwasdevelopedbasedupontheresultsofresearchbytheNishikawaLaboratory,DepartmentofMolecularGeneticsattheGraduateSchoolofMedicine,KyotoUniversity.Itisusedformolecularstudiesonproliferationandmaintenancemechanismsofmousehematopoieticstemcellsandhasbeenreportedeffectiveinthesame.

Themediumisserum-free,whicheliminateseffectsofanyunknownfactorsinserum,anditcontainsextremelylimitedproteincomponents,allowingtheactionofcytokines,etc.tobeclearlyidentified.Further,beingcomposedofknowningredients,theproductaffordsconstancyinexperimentalconditions.(SeeReferenceLiteratures:(1),(2),(3),(4),(5))

[Formulation]

1.Basalmedium(powder)

ConstitutedfromacompositeofslightlymodifiedRPMI1640,Dulbecco"sMEM,andHam"sF-12.

2.Sodiumhydrogencarbonate(powder)

3.Supplement(liquid)

HEPESbuffercontaininghormonegrowthfactors(insulin,transferrin,sodiumseleniteandethanolamine).Proteincomponentsotherthaninsulinandtransferrinhavenotbeenadded.

[DirectionsforUse]

(PreparingtheMedium)

1.Add1bottleofthebasalmedium(10.7g)toapproximately950mLofpurifiedwaterandmixwelltodissolve.

2.Tothesolutionabove,add1vial(2.2g)ofsodiumhydrogencarbonateanddissolve,andfurtherdilutetoatotal1,000mL.

3.Sterilizebyfilteringthroughalowadsorptionmembranefilterofporesize0.2-0.22um(ToyoRoshiKaishaLtd.,Cat.No.C020A047AorMilliporeCorporation,Cat.No.GVWP04700,etc.).

4.Asepticallyadd1vialofthesupplement(10mL)tothefiltered1,000mLofmedium.

5.Sealandstorerefrigerated(2-10C).

(ExamplesofUsage)

1.MethodforcultivationofBprecursorcellsonamousestromalcelllineST2layerusingSF-O3(AddIL-7:10U/mL,2-mercaptoethanol:100umol/L).

1)SeedtheST2cellsatadensityof1.5x10^5cells/welltomakethefeederlayer.

2)Seedthemousebonemarrowcellsatadensityof2x10^5cells/wellontheST2feederlayerandcontinuetoincubate,replacinghalfofthemediumevery4-5days.Wellused:FalconPrimariaCultureware6-WellPlate(3846)

2.MethodforcultivationofhematopoieticprecursorcellsinamousestromalcelllinePA6layerusingSF-O3(AddbFGForEGF:5-10ng/mL,2-mercaptoethanol:100umol/L).

1)SeedthePA6cellsatadensityof2x10^5cells/welltomakethefeederlayer.

2)Seedthemousebonemarrowcellsatadensityof2x10^5cells/wellonthePA6feederlayerandcontinuetoincubate,replacinghalfofthemediumevery4-5days.Wellused:FalconPrimariaCultureware6-WellPlate(3846)

[PrecautionsforUseandHandling]

1.Theentirebottleofbasalmediumshouldbedissolvedinasingleuse.Avoiddissolvingthepowderinsmallportion.

2.Whendissolvingthebasalmedium,injectcarbondioxidegas(high-qualitycarbondioxideordryiceissuitable)toshiftthepHofthemediumtoacidic.AnacidicpHenhancessolubilityofthemediumaswellasspeedsequilibration(pH7.2-7.4)ofthemediuminthecarbondioxidegasincubator.

3.Asthesupplementisaconcentratedsolution,itmayshowprecipitationwhenstoredrefrigerated.Itshouldbemixedwellbeforeaddingtothebasalmedium.

4.Avoidfilteringthemediumafterthesupplementhasbeenadded,asitmaycauseloweredgrowthinsometypesofcells.

5.Thepreparedmediumisconfirmedtobestablefor3monthswhenstoredat2-10C,however,itshouldbeusedassoonaspossIBLe.

6.Thisproductdoesnotcontain2-mercaptoethanol.

7.Thisproductisareagentforinvitroresearchuseonlyandshouldnotbeusedfordiagnosticpurposes,norshouldthegrowncellsbeusedfortreatment.

8.Treatmentofwastesgeneratedbythismethodshouldbedoneaccordingtoguidelinesinaccordancewiththelocallawsandregulations.

**Note)Pleasealsorefertothe"S-CloneSF-O3"MaterialSafetyDataSheet(MSDS)beforeuse.

MSDSforSF-O3Cat#1303(PDFFile)

[StorageandShelfLife]

*Storeat2-10C,avoidfreezing.

*Theshelflifeis1yearfollowingmanufacture(expirationdateisindicatedontheouterbox)

[PackageContents]

*Basalmedium:1L-equivalentx1bottle

*Sodiumhydrogencarbonate:2.2gx1vial

*Supplement:10mLx1unitvial

[ReferenceLiterature]

(1)ImmunologyLetters.40:163-169,1994

(2)SeminarsinIMMUNOLOGY,7:185-196,1995

(3)J.Exp.Med.,182:315-323,1995

(4)ClinicalImmunology,27(10):1193-1198,1995

(5)JournalofCellBIOLOGy,Volume132,Numbers1&2,January199691-99

(6)NatureBiotechnology23,1542-1550(2005)

(7)Cell138,300-313,July24,2009

(8)Blood2010116:4474-4482

(9)JCellSci122,3303-3311,September15,2009

(10)Development2013Jan1;140(1):216-25

ProductInsertSF-O3Cat#1303(PDFFile)

FORRESEARCHUSEONLY,SHOULDNOTBEUSEDFORDIAGNOSTICPURPOSE,NORSHOULDTHEGROWNCELLSBEUSEDFORTREATMENT.

Aserum-freemediumforuseinresearchofhematopoieticstemcells

Manufacturedby:SekisuiMedicalCo.,LTD