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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Cell Biologics 是一家国际领先的原代细胞和细胞培养耗材供应商，提供多领域高品质的人源或动物原代细胞、包括内皮细胞、上皮细胞、肿瘤和干细胞，并提供优化的细胞培养基和其他相关产品。Cell Biologics 还首次提供基因修饰动物来源如鼠源的原代内皮细胞，以及根据用户特殊的科研需求订制细胞。公司同时发展靶向自身免疫疾病的治疗平台和新型临床生物制剂的输送系统。

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肿瘤细胞系：人肿瘤细胞、小鼠肿瘤细胞

干细胞：大鼠间充质干细胞、干细胞培养基、小鼠骨髓祖细胞、小鼠胚胎干细胞、小鼠间充质干细胞

动物组织器官：小鼠组织和器官、小鼠体液、大鼠组织和器官、大鼠体液

分子生物学：大鼠及小鼠蛋白裂解液，RNA, DNA

其他产品（犬、仓鼠细胞）：犬原代细胞、仓鼠原代细胞、GFP表达小鼠内皮细胞