歡迎您致電 華雅再生醫(yī)學(xué)旗艦公司:紅榮微再(上海)生物工程技術(shù)有限公司 :152 1681 4001。華雅再生醫(yī)學(xué)-客服: 316 808 6348
The TeSR™ family of feeder-free media are produced using rigorously pre-screened materials to ensure the highest levels of batch-to-batch consistency and experimental reproducibility, allowing you to minimize variation in your research and establish a
continuous TeSR™ media-based workflow. Each medium is based on published formulations1-3 from the laboratory of James Thomson, and allows researchers to maintain high quality embryonic stem (ES) cell and induced pluripotent stem (iPS) cell culture systems.
Which TeSR™ Feeder-Free Medium is Right for You?
Reprogram
TeSR™-E7™ is a xeno-free reprogramming medium optimized for human iPS cell induction from fibroblasts.
Based on the E8 formulation³ with TGFβ removed for improved reprogramming efficiency.
Pre-screened components ensure generation of high quality iPS cell colony morphology for easy identification and rapid subcloning.
ReproTeSR™ is a xeno-free reprogramming medium optimized for the generation of iPS cells from blood-derived cells.
Formulation is optimized for reprogramming of erythroid or CD34+ cells expanded in vitro from peripheral blood.
Rapid emergence of large colonies with high quality iPS cell-like morphology.
歡迎您致電 華雅再生醫(yī)學(xué)旗艦公司:紅榮微再(上海)生物工程技術(shù)有限公司 :152 1681 4001。華雅再生醫(yī)學(xué)-客服: 316 808 6348
Maintain
mTeSR™1 is the most published medium for the culture of hPSCs.
Over 700 peer-reviewed publications, including 35 Nature Publishing Group and 25 Cell Press.
Maintained thousands of ES and iPS cell lines for more than 7 years in over 50 countries.
Utilized in published protocols for a wide variety of applications including bioreactor expansion, single-cell cloning, and lineage-specific differentiation of mTeSR™1-maintained hPSCs.
Contains pre-screened bovine serum albumin to stabilize the medium, aid in lipid-nutrient transport, and protect from cellular toxins and stresses.
Maintains pluripotency of ES and iPS cells after extended periods in culture and supports the derivation of iPS cells and their in vitro differentiation into mature cell types.
TeSR™2 is a xeno-free version of mTeSR™1.
• Modified formulation, but with all xeno-free components, to produce a more defined medium.
• Compatible with published mTeSR™1 protocols for a wide variety of applications.
• Contains recombinant human albumin to aid in lipid/nutrient transport and protect cultures from
cellular toxins and stresses.
TeSR™-E8™ is a xeno-free culture medium, based on the published E8 formulation.3,4
• Contains only the 8 most essential components required for PSC maintenance.
• Has 433x less protein than mTeSR™1.
• Can be used with Corning® Matrigel® hESC-qualified matrix, or for a compley xeno-free system, with Vitronectin XF™.
Differentiate
Lineage-Specific Differentiation with STEMdiff™
Consistent differentiation of ES and iPS cell lines to downstream lineages can be challenging. We recommend our STEMdiff™ suite of products for optimal and reproducible differentiation to cells of the:
• Ectodermal Lineage: STEMdiff™ Neural Induction Medium, STEMdiff™ Neural Progenitor Medium, and STEMdiff™ Neural Progenitor Freezing Medium
• Endodermal Lineage: STEMdiff™ Definitive Endoderm Kit and STEMdiff™ Definitive Endoderm Kit (TeSR™-E8™-Optimized)
• Mesodermal Lineage: STEMdiff™ APEL™ Medium and STEMdiff™ APEL™-LI Medium
Low-Protein Differentiation with TeSR™-E5 and TeSR™-E6
TeSR™-E6 and TeSR™-E5 are low protein media that can be used for differentiation, screening assays and other applications where the removal of specific cytokines is desirable. Other cytokines and growth factors can be added to these media for specific applications.
• TeSR™-E6 is a defined, serum- and xeno-free medium that is based on the formulation of TeSR™-E8™ but does not contain bFGF, TGFβ.³
• TeSR™-E5 is also a defined, serum- and xeno-free medium based on the formulation of TeSR™-E8™ but does not contain bFGF or TGFβ or insulin.³
歡迎您致電 華雅再生醫(yī)學(xué)旗艦公司:紅榮微再(上海)生物工程技術(shù)有限公司 :152 1681 4001。華雅再生醫(yī)學(xué)-客服: 316 808 6348
Cryopreserve
mFreSR™ and FreSR™-S are defined, serum-free media optimized for cryopreservation of ES or iPS cells cultured in TeSR™ maintenance media. hPSCs cryopreserved in FreSR™ have higher thawing efficiencies than those reported with conventional methods using serum.5-8
• Use mFreSR™ for cryopreservation of human ES and iPS cells as cell aggregates.
• Use animal component-free FreSR™-S for cryopreservation of cells in single cell suspension.
Corning Matrigel Basement Membrane Matrix基底膜基質(zhì)膠(標(biāo)準(zhǔn)型) 5mL/瓶 華雅再生醫(yī)學(xué) BD 356234 ¥2,023
Corning Matrigel Basement Membrane Matrix基底膜基質(zhì)膠(標(biāo)準(zhǔn)型) 10mL/瓶 華雅再生醫(yī)學(xué) BD 354234 ¥3,859
Corning Matrigel Basement Membrane Matrix基底膜基質(zhì)膠(高濃度) 10mL/瓶 華雅再生醫(yī)學(xué) BD 354248 ¥6,045
Corning Matrigel Matrix基底膜基質(zhì)膠,低生長(zhǎng)因子GFR(標(biāo)準(zhǔn)型) 5mL/瓶 華雅再生醫(yī)學(xué) BD 356230 ¥2,325
Corning Matrigel Matrix基底膜基質(zhì)膠,低生長(zhǎng)因子GFR(標(biāo)準(zhǔn)型) 10mL/瓶 華雅再生醫(yī)學(xué) BD 354230 ¥4,533
Corning Matrigel Matrix基底膜基質(zhì)膠,低生長(zhǎng)因子GFR,無(wú)酚紅(標(biāo)準(zhǔn)型) 10mL/瓶 華雅再生醫(yī)學(xué) BD 356231 ¥4,883
Corning Matrigel Invasion Chamber,8.0um PET膜,24個(gè)包被小室 24個(gè)/箱 華雅再生醫(yī)學(xué) BD 354480 ¥4,301
加拿大的STEMCELL公司(STEMCELL Technologies Inc.)近日宣布將推出一系列新產(chǎn)品,以加速人多能干細(xì)胞的研究和轉(zhuǎn)化應(yīng)用。這些新產(chǎn)品包括全新的多能干細(xì)胞培養(yǎng)基TeSR-E8,以及誘導(dǎo)多能干細(xì)胞定向分化的STEMdiff系列產(chǎn)品。
加拿大的STEMCELL公司(STEMCELL Technologies Inc.)近日宣布將推出一系列新產(chǎn)品,以加速人多能干細(xì)胞的研究和轉(zhuǎn)化應(yīng)用。這些新產(chǎn)品包括全新的多能干細(xì)胞培養(yǎng)基TeSR-E8,以及誘導(dǎo)多能干細(xì)胞定向分化的STEMdiff系列產(chǎn)品。
TeSR™-E8™是一種成分高度確定、無(wú)需滋養(yǎng)層細(xì)胞(feeder-free)的培養(yǎng)基,適用于人胚胎干細(xì)胞(hESC)和人誘導(dǎo)多能干細(xì)胞(iPSC)。TeSR-E8僅包含維持hESC和iPSC所需的zui關(guān)鍵成分,為多能干細(xì)胞的培養(yǎng)提供了一種更簡(jiǎn)單的條件。
此培養(yǎng)基是基于James Thomson實(shí)驗(yàn)室去年發(fā)表在《Nature Methods》上的E8配方。作為iPSC創(chuàng)始人之一,美國(guó)威斯康辛大學(xué)麥迪遜分校的James Thomson教授領(lǐng)導(dǎo)的研究小組曾開發(fā)出mTeSR™1和TeSR™2培養(yǎng)基。盡管TeSR培養(yǎng)基能夠在沒有動(dòng)物蛋白的情況下支持干細(xì)胞的生長(zhǎng),但其中添加了人血清白蛋白和人源基質(zhì)蛋白,這就使得培養(yǎng)條件極其昂貴,不適合日常使用。
James Thomson教授領(lǐng)導(dǎo)的研究小組對(duì)iPSC的培養(yǎng)條件進(jìn)行了優(yōu)化,去除了培養(yǎng)基中的BSA和BME,并在DMEM/F12培養(yǎng)基中添加了胰島素、FGF2、維生素C、硒、轉(zhuǎn)鐵蛋白和TGFβ,開發(fā)出一種*確定的培養(yǎng)基E8。這種培養(yǎng)基能夠支持人ESC和iPSC的未分化增殖。
TeSR-E8可與BD Matrigel™ hESC基質(zhì)或玻璃粘連蛋白(vitronectin)共同使用。根據(jù)STEMCELL與威斯康辛大學(xué)校友研究基金會(huì)的協(xié)議,TeSR-E8培養(yǎng)基預(yù)計(jì)在今年9月份上市。
同時(shí),STEMCELL還推出了STEMdiff系列產(chǎn)品,包括STEMdiff™ Definitive Endoderm / Cardiomyocyte / Hematopoietic kits。這些新的試劑盒可誘導(dǎo)hESC/iPSC定向分化成內(nèi)胚層、心肌細(xì)胞和造血祖細(xì)胞。STEMdiff產(chǎn)品采用可靠的試劑和操作步驟,可使多能干細(xì)胞穩(wěn)定地向多種譜系細(xì)胞分化。此產(chǎn)品經(jīng)過(guò)優(yōu)化,可與mTeSR™1和TeSR™2培養(yǎng)基共同使用,預(yù)計(jì)在今年10月份上市。
有了TeSR-E8和新的STEMdiff產(chǎn)品線,STEMCELL繼續(xù)向人類多能干細(xì)胞研究提供全面、成分確定的完整工具。這些高質(zhì)量的試劑將提高實(shí)驗(yàn)重復(fù)性,避免培養(yǎng)基中的可變成分,并增加ES細(xì)胞和iPS細(xì)胞培養(yǎng)的臨床相關(guān)性。
歡迎您致電 華雅再生醫(yī)學(xué)旗艦公司:紅榮微再(上海)生物工程技術(shù)有限公司 :152 1681 4001。華雅再生醫(yī)學(xué)-客服: 316 808 6348
BeYaMATM 1 Juvi Human iPSC/ESC Growth Medium
人多能干細(xì)胞無(wú)血清培養(yǎng)液*實(shí)驗(yàn)操作手冊(cè)
一、 培養(yǎng)皿包被
1、 4℃溶解Matrigel(BD 354277)八小時(shí),輕搖混勻。請(qǐng)注意:原液應(yīng)是均質(zhì)粘稠的半流體狀態(tài),要避免溫度過(guò)高,否則Matrigel會(huì)變成凝膠狀態(tài)。
2、 冰上預(yù)冷1.5ml離心管和移液器槍頭,冰上快速將Matrigel分裝成350-500ul/支,儲(chǔ)存于-80℃,用前置于4℃解凍。
3、 將溶解的Matrigel加入25ml預(yù)冷的無(wú)血清DMEM/F12培養(yǎng)基中,輕輕混勻,避免產(chǎn)生氣泡。
4、 將稀釋的Matrigel加入六孔板中,1ml/孔,輕搖培養(yǎng)板,使Matrigel均勻鋪在培養(yǎng)板底部。
5、 Parafilm封好包被的培養(yǎng)板,至于4℃儲(chǔ)存,用前置于37℃培養(yǎng)箱中孵育至少1小時(shí)。
二、 培養(yǎng)液準(zhǔn)備
1、 在4℃解凍BeYaMATM 1 5X Supplement(貨號(hào):HJ0103M)添加劑,加入BeYaMATM 1 Basal Medium(貨號(hào):HJ0102M)基礎(chǔ)培養(yǎng)基中,形成500mL BeYaMATM 1 Complete Medium(貨號(hào):HJ0101M)*培養(yǎng)基。
2、 BeYaMATM 1*培養(yǎng)基可在4℃穩(wěn)定儲(chǔ)存2周。可按實(shí)際用量將BeYaMATM 1*培養(yǎng)基分裝后置于-80或-20℃保存,避免反復(fù)凍融。
三、 hESCs/hiPSCs復(fù)蘇
1、 復(fù)蘇hESCs/hiPSCs前,復(fù)溫Matrigel包被的培養(yǎng)板,吸棄孔內(nèi)液體,加入2ml BeYaMATM 1*培養(yǎng)液及2ul 5uM/L Rock Inhibitor Y27632,用以促進(jìn)細(xì)胞貼壁。
2、 從液氮罐中取出hESCs/hiPSCs凍存管,浸入37℃水浴,輕輕搖晃,直到細(xì)胞融化至僅剩一小塊冰晶,迅速取出并用75%酒精擦拭凍存管外表面,轉(zhuǎn)移至無(wú)菌超凈臺(tái)中。
3、 將細(xì)胞懸液接種至準(zhǔn)備好的培養(yǎng)板中,輕輕晃動(dòng)使細(xì)胞均勻分布在板底。
4、 小心將培養(yǎng)板放置于培養(yǎng)箱中,避免振動(dòng)。
5、 30-90min后顯微鏡下觀察細(xì)胞基本貼壁,更換新鮮BeYaMATM 1*培養(yǎng)液,之后每24h換液,4-6天后傳代。
四、 hESCs/hiPSCs傳代
1、 提前準(zhǔn)備好Matrigel(BD 354277)包被過(guò)的六孔板,吸棄孔內(nèi)包被液,加入2ml BeYaMATM 1*培養(yǎng)液。
2、 吸棄待傳代hESCs/hiPSCs孔內(nèi)的培養(yǎng)液,并用1ml無(wú)鈣鎂的PBS洗1遍。
3、 加入0.5ml Accutase細(xì)胞分離液使之*覆蓋皿底。
4、 室溫孵育5-8分鐘或37℃孵育3-5分鐘,顯微鏡下觀察到大部分克隆細(xì)胞間出現(xiàn)間隙,細(xì)胞表面發(fā)亮但尚未相互分離時(shí),可吸去Accutase。
5、 可將培養(yǎng)板放置37℃繼續(xù)孵育1min,或者直接加入適量新鮮BeYaMATM 1*培養(yǎng)液,輕輕搖晃,干細(xì)胞集落基本脫落。亦可用細(xì)胞刮刀將剩余克隆輕輕刮下,盡量避免用槍頭反復(fù)吹打細(xì)胞。
6、 將制備好的細(xì)胞懸液按1:3 - 1:6的比例接種至準(zhǔn)備好的培養(yǎng)板中,注意細(xì)胞過(guò)密則克隆邊界不圓滑,易分化,細(xì)胞過(guò)稀則克隆存活率降低。
7、 輕輕搖晃培養(yǎng)板讓細(xì)胞均勻分散,置于37℃,5% CO2培養(yǎng)箱中培養(yǎng)8小時(shí)。
8、 每天更換培養(yǎng)液,待細(xì)胞密度達(dá)到80%以上,或者克隆中央密度過(guò)大,影響細(xì)胞生長(zhǎng)時(shí)進(jìn)行傳代。
如果您在使用BeYaMATM 1 JuviTM Human iPSC/ESC Growth Medium 人多能干細(xì)胞無(wú)血清培養(yǎng)液的過(guò)程中,有任何技術(shù)問題,我們開通了Juvi科學(xué)家技術(shù)團(tuán)隊(duì),歡迎您發(fā)郵件到Juvi@hystemcell.com 進(jìn)行交流溝通,我們的目標(biāo)是:推動(dòng)中國(guó)iPS干細(xì)胞技術(shù)走向產(chǎn)業(yè)和臨床!