RatPrimaryLungMesenchymalStemCells
CatalogNo.R2303
SuggestedMedium: M5566–MesenchymalStemCellMedium/wKit-500ml
ProductDescription
RatLungPrimaryMesenchymalStemCellsarederivedfromthelungtissuesofday-1Sprague-DawleyRats.RatLungPrimaryMesenchymalStemCellsaregrowninT25tissuecultureflaskspre-coatedwithgelatin-basedsolutionfor0.5hourandincubatedinCellBIOLOGics’CellCultureMediumfor7-15days.Culturesarethenexpanded.Priortoshipping,cellsaredetachedfromflasksandimmediatelycryo-preservedinvials.Eachvialcontainsatleast1x106cellspermlandisdeliveredfrozen.RatLungPrimaryMesenchymalStemCellsarenegativeforbacteria,yeast,fungi,andmycoplasma.Cellscanbeexpanded3-6passagesbytheratioof1:2underthecellcultureconditionsspecifiedbyCellBiologics.Repeatedfreezingandthawingofcellsisnotrecommended.MouseLungMesenchymalStemCellsaretestedforexpressionofMarkersusingantibodies,CD29andCD44positivebyflowcytometry.
LaboratoryApplications
RatLungPrimaryMesenchymalStemCellscanbeusedinassaysofstandardbiochemicalproceduresperformedwithcellculturesincludeRT-PCR,Westernblotting,immunoprecipitation,orimmunofluorescentflowcytometry.
StorageofCellBiologicsProducts
CellBiologicsshipsfrozencellsondryice.Onreceipt,immediatelytransferfrozencellstoliquidnitrogen(-180°C)untilreadyforexperimentaluse.Live-cellshipmentisalsoavailableonrequest.
Nevercanprimarycellsbekeptat-20°C.
AuthorizedUsesofCellBiologicsProducts
RatLungPrimaryMesenchymalStemCellsfromCellBiologicsaredistributedforresearchpurposesonly.Ourproductsarenotauthorizedforhumanuse,forinvitrodiagnosticprocedures,orfortherapeuticprocedures.TransferorresaleofanyCellBiologics’cellsorproductsfromthepurchasertoothermarkets,organizationsorindividualsisprohibitedbyCellBiologicswithoutthecompany’swrittenconsent. CellBiologics’TermsandConditionsmustbeacceptedbeforesubmittinganorder.
Disclaimer
AlthoughRatLungPrimaryMesenchymalStemCellsareisolatedfromlaboratorymicetestingpathogen-free,investigatorsshouldhandlethecellsthattheyreceivefromCellBiologicswithcaution,sincenotestprocedurecancompletelyguaranteetheabsenceofinfectiousagents.
WarrantyandLiABIlity
CELLBIOLOGICS’guaranteeappliesonlytoyourpurchaseofCELLBIOLOGICS'cellswithCELLBIOLOGICS’MediaandCoatingSolutionforappropriatecellcultureandcelltestingfollowingCELLBIOLOGICS’onlineprotocolswithin35daysfromthedateofproductdelivery.
GeneralProtocolforRecoveringorFreezingPrimaryCells
Allcellcultureproceduresmustbeconductedinabio-safetycabinet.
Anyandallmedia,supplements,andreagentsmustbesterilizedbyfiltrationthrougha0.2µmfilter.
Useaseptictechniquetopreventmicrobialcontamination.
Cryo-preservedcellsmustbestoredinliquidnitrogenorseededimmediatelyuponarrival.
Medium:
ReviewtheinformationprovidedontheCellBiologics’websiteaboutappropriateculturemedia(e.g.serumandothersupplements).Usepre-warmed(37°C)cellculturemedia(30-50ML)torecovercryo-preservedcellsandwhenchangingmediaorsplittingcells.
Coatingofflasksordishes:
CoatsterileculturedishesorflaskswithGelatin-BasedCoatingSolution(CellBiologics,CatalogNo.6950)for10-30minandthenaspiratetheexcesssolutionbeforeseedingcells.
Cellrecoveryfromcryovial:
• Quicklythawcellsincryo-vialbyincubatingthemina37°Cwaterbathfor<1minuntilthereisjustasmallbitoficeleftinthevial.
• Promptlyremovethevialandwipeitdownwith70%ethanol.
• Transfercellsfromthevialtoasterilecentrifugetube.Add8-10mlofpre-warmedCellBiologicsCellCultureMedium.
• Flushthevialwithanadditional0.5-1mlofmediumtoensurecompletetransferofcellstothecentrifugetube.
• Centrifugecellsat200gfor5minutes.
• AspiratethesupernatantandresUSPendthecellpelletin6mlofCellBiologics’CellCultureGrowthMedium.
• AddresuspendedcellsintoaT75flaskpre-coatedwithGelatin-BasedCoatingSolution(CellBiologics,CatalogNo.6950).
• PlacetheT75flaskinahumidified,5%-CO2incubatorat37°C.
• Changeculturemediathefollowingdaytoremovenon-adherentcellsandreplenishnutrients.
• Changecellculturemediumeverydaywhencellsare>70%confluent.
• Cellsshouldbecheckeddailyunderamicroscopetoverifyappropriatecellmorphology.
Expansionofculturedprimarycells:
• Removeanddiscardthecellculturemediafromtheflask.
• Flushtheadherentlayer2timesusinga5mlsterilePipettewithsterilePBS(1X)withoutcalciumandmagnesiumtodislodgelooselyattachedcellsandremovefraction.
• Removeanddiscardthewashsolutionfromtheflask.
• Incubatecellswithwarm(37°C)0.05%Trypsin-EDTAsolution(CellBiologics,CatalogNo.6915)for2-5minutes.Use3.0mlofTrypsin-EDTAsolutionwhencollectingcellsfromT75flasks,and2mlwhenusingT25flasks.Assoonascellshavedetached(theflaskmayrequireafewfirmgentletaps),add8-10mlofCellBiologics’CellCultureMediumsupplementedwith5-10%FBStoaT25orT75flask(theFBSwillneutralizethetrypsin).
• PlatecellsinfreshflasksorplatesprecoatedwithGelatin-BasedCoatingSolutioninahumidified,5%-CO2incubatorat37°C.
• Changeculturemediathefollowingdaytoremovenon-adherentcellsandreplenishnutrients.
• Cellsshouldbecheckeddailyunderamicroscopytoverifyappropriatecellmorphology.
• Changeculturemediumevery24-48hours.Pleasenotethatthemediumshouldbechangedeverydaywhencellsare>70%confluenttoremovenon-adherentcellsandreplenishnutrients.Pre-washcellswith1XPBS1-2timeswheneverreplacingthemedium.
Werecommendsplittingprimarycellsatthefollowratio:
• Therecommendedsplitratioforprimarymurinecellsis1:2or1:3.
ProcedureforFreezingCells
Materials:
• 1XPhosphateBufferedSaline(PBS-1X)
• 0.05%Trypsin-EDTA(1X)solution(CellBiologics,CatalogNo.6915)
• TissueCultureMedia
• ColdFreezingMedia(10%DMSO,20%FBSand70%culturemedium,CellBiologics,CatalogNo.6916).
• LabeledCryovials
• Confluentcells
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Chapter 3 CELL CULTURE MEDIA 培养基 mECM1166 小鼠内皮细胞培养基 Mouse Endothelial Cell Growth Medium /w BulletKit 500 ml mECM1168 小鼠内皮细胞培养基 Mouse Endothelial Cell Growth Medium /w BulletKit 500 ml mECM1169 小鼠内皮细胞培养基 Mouse Endothelial Cell Growth Medium /w BulletKit 250 ml mFBM2267 小鼠成纤维细胞培养基 Mouse Fibroblast Growth Medium /w BulletKit 500 ml mBMM3366 小鼠骨髓细胞培养基 Mouse Bone Marrow Leukocyte Culture Medium /w BulletKit 500 ml mBMM3367 小鼠骨髓中性粒细胞培养基 Mouse Bone Marrow Neutrophil Medium /w BulletKit 500 ml mBMM3368 小鼠骨髓巨噬细胞培养基 Mouse Bone Marrow Macrophage Culture Medium /w BulletKit 500 ml mKSM5566 小鼠骨髓间充质干细胞培养基 Mouse Bone Marrow Mesenchymal Stem Cell Medium /w BulletKit 500 ml mBMM3369 小鼠骨髓干细胞培养基 Mouse Bone Marrow Progenitor Cell Medium /w BulletKit 500 ml mKSM5567 小鼠细胞培养基 Mouse Cell Growth Medium /w BulletKit 500ml mEPM6621 小鼠上皮细胞培养基 Mouse Epithelial Cell Growth Medium /w BulletKit 500 ml mEPM6622 小鼠上皮细胞培养基 Mouse Epithelial Cell Growth Medium /w BulletKit 500 ml mEPM6623 小鼠上皮细胞培养基 Mouse Epithelial Cell Growth Medium /w BulletKit 250 ml mDCM7711 小鼠骨髓树突细胞培养基 Mouse Bone marrow-dendritic cells Medium /w BulletKit 500 ml mESCM8811 小鼠胚胎干细胞培养基 Mouse Embryonic Stem Cell Medium /w BulletKit 500 ml Chapter 4 CELL GROWTH SUPPLEMENT 1166 内皮细胞生长助生长溶液-ECGS Endothelial Cell Growth Supplement-ECGS 5 ml 1167 内皮细胞助生长溶液-V Endothelial Cell Growth Supplement-V 5 ml 2267 小鼠成纤维细胞生长因子 Mouse Fibroblast Growth Supplement 5 ml 3368 小鼠骨髓巨噬细胞助生长溶液 Mouse Bone Marrow Macrophage Culture Supplement 5ml 3369 小鼠骨髓干细胞助生长溶液 Mouse Bone Marrow Macrophage Growth Supplement 5ml 5566 小鼠骨髓间充质干细胞助生长溶液 Mouse Bone Marrow Mesenchymal Stem Cell Growth Supplement 5 ml 6621 小鼠上皮细胞助生长溶液-I Mouse Epithelial Cell Growth Supplement-ITS 5 ml 6622 小鼠上皮细胞助生长溶液-E Mouse Epithelial Cell Growth Supplement-EGF 5 ml 6911 胎牛血清-2 ml Fetal Bovine Serum (FBS) 2 ml 6912 胎牛血清-20 ml Fetal Bovine Serum (FBS) 25 ml 6913 胰酶/EDTA消化液 (T/E)-50 ml 0.05% Trypsin/EDTA Solution 50 ml 6914 胰酶/EDTA消化液 (T/E)-100 ml 0.05% Trypsin/EDTA Solution 100 ml 6915 非酶细胞裂解液 Enzyme-free Cell Dissociation solution 100 ml 6916 细胞冻存液 Cell Freezing Medium 50 ml 6917 青霉素/链霉素溶液-5 ml Penicillin/Streptomycin Solution 5 ml 6918 青霉素/链霉素溶液-25 ml Penicillin/Streptomycin Solution 25 ml 6919 抗霉菌溶液-25 ml Antibiotic/Antimycotic Solution 25 ml 6920 抗霉菌溶液-5 ml Antibiotic/Antimycotic Solution 5 ml 6921 左旋谷氨酰胺溶液 L-Glutamine-200 mM (100X), liquid 5 ml 6922 非必需氨基酸溶液 Non-Essential Amino Acids Solution 10 mM (100X), liquid 5 ml 6923 丙酮酸钠溶液 Sodium Pyruvate Solution 100 mM (100X), liquid 5 ml 6924 维生素溶液 Vitamin Solution (100X), liquid 5 ml 6925 2-巯基乙醇溶液 2-Mercaptoethanol (1,000X), liquid prepared in D-PBS， 5 ml 6926 胰岛素铁硒传递蛋白 Insulin-Transferrin-Selenium (100X) 5 ml 6927 牛脑垂体提取物 Bovine Pituitary Extract 25 ml 6928 HEPES 缓冲液 HEPES Buffer Solution (1 M) 5 ml 6929 胰酶/EDTA消化液 (T/E)-50 ml 0.25% Trypsin/EDTA Solution 50 ml
