Alstembio蚂蚁淘在线

商品

品牌

文章

热门搜索：

快报

蚂蚁淘特色产品FLOWTECH造影剂蚂蚁淘授权代理品牌-德国Emfret 蚂蚁淘旗下188精品现货平台正式上线啦！！！蚂蚁淘四月促销蚂蚁淘获得vitanavitech代理商授权蚂蚁淘获得QA-bio代理商授权蚂蚁淘获得Viagenbiotech代理商授权 polysciences公司24765-1到现货了(内附代理商整理说明书）蚂蚁淘四月团建“环苏州古城一周” Clodronate Liposomes用量更正！

品牌首页品牌新闻品牌简介产品中心品牌咨询联系我们

品牌分类

品牌咨询

联系方式

公司地址

苏州工业园区生物纳米园A4#216

联系电话

4000-520-616 / 18915418616

传真号码

0512-67156496

电子邮箱

info@ebiomall.com

公司网址

https://www.ebiomall.com

蚂蚁淘在线 / 品牌中心 / Alstembio / Alstembio/HumanneuralStemcelline/hNSC11

Alstembio/HumanneuralStemcelline/hNSC11

价格

￥13900.00

货号：hNSC11

浏览量：127

品牌：Alstembio

服务

全国联保

正品保证

正规发票

签订合同

加入收藏

立即购买

商品描述

NeuralStemCellLine,Human:#hNSC11

ProductDescription

Neuralstemcells(NSCs)areneuralProgenitorcellsthatarecapableofdifferentiatingtodifferentkindsofcellsinthenervoussystemwhenunderdefinedconditions.Takingadvantageoftheirconsistentpropagation,NSCshavegreatpotentialforthestudyofneurogenesis,neurodegenerativediseases,andclinicaltransplantationapplications.Alstem"shumanNeuralStemCellLineisderivedfromhumaninducedpluripotentstemcells,whicharegeneratedfromadultskinfibroblastsusingAlstem’suniquefootprint-freeepisomaliPSCreprogrammingmethod.ThesecellsareabletoproliferateregularlywithoutdifferentiatingtoneurallineagecellswhenculturedinbFGFandEGFcontainingmedium.

NeuronsderivedfromAlstem"sNSCsstainedwithbeta-III-tubulin

hNSCFeatures

Derivedfromfootprint-freehumaniPScells
Homogeneouspopulation
Candifferentiatetoover90%ofneuronsunderdefinedconditions

ProductSpecifications
ProductName	HumanNeuralStemCellLine
Catalog#	hNSC11
Size	1x10⁶cells/vial
Shipping	DryIce
StorageandStABIlity	Storeingasphaseofliquidnitrogenimmediatelyuponreceipt.Thisproductisstablefor6monthswhenstoredasdirected.
QualityControl	EachlotofhNSCsistestedforgrowthandviabilityafterrecoveryfromcryopreservation.Inaddition,eachlotistestedforexpressionofNestintoensureitsundifferentiatingcharacteristics.
SafetyPrecaution	ALSTEMhighlyrecommendsthatprotectivegloves,alabcoat,andafull-facemaskalwaysbewornwhenhandlingfrozenvials.Itisimportanttonotethatsomeliquidnitrogencanleakintothevialswhensubmersedinliquidnitrogen.Uponthawing,theliquidnitrogenreturnstothegasphase,resultinginexcessivepressurewithinthevialthatcancausethevialtoexplodeorexpelthecapwithdangerousforce.
RestrictedUse	ForResearchUseOnly.Notforuseindiagnosticortherapeuticprocedures.
Protocol	Download

Overview
Procedure

Thisprotocolcanbeusedforculturinghumanneuralstemcells.TheneuralstemcellsarederivedfromALSTEM"sfootprint-freeiPScellsandeachlotistestedfortheexpressionofNestintoensureitsundifferentiatingcharacteristics.

Protocol

Preparationofneuralstemcellculturemedium

1.ThawN-2supplement(100X)(Cat.no.17502-048,LifeTechnologies)andB-27serumfreesupplement(50X)(Cat.no.17504-044)overnightat4°C.

2.Addthe5mLofthawedN-2supplement,10mLofthawedB-27supplement,20ng/mlbFGF(Cat.no.bFGF-50,StemRD)and20ng/mlEGF(Cat.no.EGF-50,StemRD)to500mLofDMEM/F12medium(Cat.no.15-090-CV,Cellgro)aseptically,andmixwell.Filterthrougha0.2μm,low-proteinbindingfilter,ifdesired.

3.Aliquotintoappropriateamountaccordingtousageandstorethealiquotsat4°C.

Coatingplateswithpoly-l-ornithineandlaminin

1.Dilutepoly-l-ornithine(Cat.no.P3655,Sigma)oniceusingsteriletissueculture-gradewater.Aliquotthedilutedpoly-l-ornithinandkeepthealiquotsin-80°Cuntiluse

2.Aliquotlaminin(Cat.no.L2020,Sigma)oniceandkeepthealiquotsin-80°Cuntiluse.

3.Add1mLofpoly-l-ornithine(20µg/mL)toonewellofa6-wellplate,andincubateat37°Cfortwohours.

4.Aspiratepoly-l-ornithineandrinsethewelltwicewith1mLofDPBS.Add1mLoflamininsolution(5µg/mL)inthewellandincubateat37°Cfor2hours.

5.Aspiratethelamininbeforeplatingcellsorstoreplateat4°Cuntilneeded.

CoatingplateswithMatrigel(alternativeplatecoatingmethod)

Matrigel(Cat.no.354277,BD)shouldbealiquotedandstoredat-80°Cforlong-termuse.

1.Thawthematrigeloniceuntilliquid.Dilutematrigel1:50withpre-chilledKODMEM/F12.

2.Immediatelyusethedilutedmatrigelsolutiontocoattissueculture-treatedplates.Fora6-wellplate,use0.8mLofdilutedmatrigelsolutioninoneofthewells,andswirltheplatetospreadthematrigelsolutionevenlyacrossthesurface.

3.Keepthecoatedplateat37°Cfor1hourorovernightat4°C.Ifplatehasbeenstoredat4°C,allowtheplatetoincubateat37°Cforatleast30minutesbeforeremovingthematrigelsolution.

Preparationofneuralstemcellfreezingmedium

1.Add9mLofneuralstemcellculturemediumwith1mLofDMSOina15mLconicaltube.Keepitat4°Cuntiluse.Prepareneuralstemcellfreezingmediumimmediatelybeforeuse.Donotstore.

ThawingcryopreservedhNSC

1.QuicklythawtheNSCsina37°Cwaterbathbygentlyshakingthecryovialcontinuouslyuntilhalfthawed.Removethecryovialfromthewaterbathandspraywith70%ethanoltosterilize.

2.Transferthecontentsofthecryovialtoa15ofmLconicaltube.Add5mLofwarmN2/B27mediumcontainingbFGFandEGFtothetube,gentlymixingasthemediumisadded.

3.Centrifugecellsat200xgfor5minutesatroomtemperature.

4.Whilecentrifuging,removethematrigelsolutionfromacoatedtissueculture6-wellplate.Add1mLofwarmN2/B27mediumcontainingbFGFandEGFtoonewellofthe6-wellplate.

5.Aftercentrifugation,aspiratethemediumfrom15mLtube.GentlyresUSPendthecellpelletin1mLN2/B27mediumcontainingbFGFandEGF,followingbytransferringthecellstothematrigelcoated6-wellplate.

6.Movetheplateinquicksidetoside,forwardtobackmotionstoevenlydistributethecellswithinthewell,andthenplacetheplateintothe37°Cincubator.Culturethecellsat37°C,with5%CO2and95%humidity.

7.Changemediumineverytwodays.

1.Passagecellswhenthecellsreach80-90%confluence.

2.AspiratemediumfromthehNSCcultureandrinsewithDPBS(2mL/well).

3.Add0.5mLperwellofaccutase(Cat.no.SCR005,Millipore).Keeptheplateat37°Cfor2-3minutes.

4.Neutralizetheaccutasewith1mlofN2/B27mediumcontainingbFGFandEGF.Gentlyrinseeachwell2-3timestoliftupmostofthehNSCs.

5.Transferthedetachedcellstoa15mLconicaltubeandrinsethewellwithanadditional2mLofN2/B27containingbFGFandEGFtocollectanyremainingcells.Addtherinsetothe15mLtube.

6.Centrifugethe15mLconicaltubecontainingthecellsat200xgfor5minutesatroomtemperature.

7.Aspiratethesupernatant.ResuspendthepelletinN2/B27containingbFGFandEGFbygentlypipettingupanddown.

8.PlatethehNSCsusing1:6ratioswith2mLN2/B27mediumcontainingbFGFandEGFinanewplatecoatedwithpoly-l-ornithineandlamininormatrigel(Removethecoatingsolutionbeforeplatingthecells).

9.Movetheplateinquicksidetoside,forwardtobackmotionstoevenlydistributethecellswithinthewells,andthenplacetheplateintothe37°Cincubator.Culturethecellsat37°C,with5%CO2and95%humidity.

10.Changemediumoncepertwodays.

CryopreservinghNSCs

1.PreparethehNSCsfreezingmediumandkeepthefreezingmediumin4°Cuntiluse.

2.Performsteps1-6fromPassagingthehNSCsasabove

3.Gentlyaspiratethesupernatantandloosenthecellpelletbytappingthebottomofthetube.

4.Gentlyresuspendthepelletinfreezingmedium.

5.Transfer1mLofcellswithfreezingmediumintoeachlabeledcryogenicvial.

6.Placevialsintoanisopropanolfreezingcontainerandplacethecontainerat-80°Covernight.

7.Transferthevialtoaliquidnitrogentankthenextday.

Alstembio定制病毒包装慢病毒包装请求（Web表单）逆转录病毒打包请求（Web表单）我们提供快速的慢病毒和逆转录病毒包装服务，以使用您的病毒构建体生产高质量，高滴度的病毒颗粒，周转时间为1-2周。节省您自己准备病毒的时间和精力，并获得随时可以转导的病毒颗粒。我们的包装服务的特点：周转时间： 10个工作日或更少质量：符合国家卫生研究院《生物安全2级》标准的一流BSL-2设备中的病毒生产，并具有严格的质量控制滴度测量：通过qPCR确定的准确病毒滴度，以测量每毫升感染单位（IFU / ml）所需材料：如果您打算准备自己的DNA，我们要求您提供10-400 µg DNA，具体取决于所需的滴度和体积可交付成果：对于100 µl的体积，我们提供5 x 20 µl的病毒颗粒等分试样，对于200 µl的体积，我们提供10 x 20 µl的等分试样。也可以使用其他配置。

资质认证

获得国家资质，权威认证！
全国联保

全国联保，官方无忧售后
正规发票

正规发票，放心购买
签订合同

签订合同，保障您的权益

/**/