人造血干細胞體外擴增試劑:StemRegenin-1(SR1)(造血干細胞擴增劑) Datasheet of StemRegenin 1Catalog: DC5142 Purity of current batch: >98%
用120nM的EC50評估方法,在5到7天之后,StemRegenin 1能增加CD34+陽性細胞數(shù)量。參照對比的質(zhì)控細胞,在mPB CD34+陽性細胞中添加細胞因子和StemRegenin 1(SR1-1μM)培養(yǎng)7天,表達CD34+, CD133+, and CD90+ 的造血干細胞和祖細胞群體數(shù)量分別增加2.6倍,2.3倍和10倍。繼續(xù)添加StemRegenin 1培養(yǎng)3周時間,總有核細胞(TNC)數(shù)量增加了11倍,表達CD34+陽性細胞數(shù)量增加了73倍,根據(jù)輸入細胞增長了1118倍。SR1(1μM)處理促進CD34+細胞增殖以及降低了VentX在人CD34+陽性細胞中的表達水平。VentX的異位表達可防止SR1誘導(dǎo)CD34+陽性細胞的膨脹。 序貫共培養(yǎng)與骨形態(tài)發(fā)生蛋白4 BMP-4(20納克/毫升),PGE2(2μM),和SR1(0.75μM)導(dǎo)致強大的豚尾猴iPSC的造血祖細胞的形成。在表達CD34的細胞群體中可以分離到CD34(+)CD38(-)Thy1(+)CD45RA(-)CD49f(+)的細胞,在添加了StemRegenin 1(SR1 0.75μM)的細胞培養(yǎng)中,數(shù)量擴大了3倍并保持著這種長期重建造血干細胞表型。 人造血干細胞體外擴增試劑:StemRegenin-1(SR1)(造血干細胞擴增劑)Description:StemRegenin 1 increases the number of CD34+ cells after 5 to 7 days with an EC50 of 120 nM. Culture of mPB CD34+ cells with cytokines plus SR1 (1 μM) for 7 days increases the number of CD34+, CD133+, and CD90+ hematopoietic stem and progenitor cell populations 2.6-, 2.3-, and 10-fold, respectively compared to control cells. Continued culture with SR1 (1 μM) for 3 weeks leads to an 11-fold increase in total nucleated cells (TNC), a 73-fold increase in CD34+ cells as compared to control cultures, and a 1118-fold increase in CD34+ cells relative to input cells. Culture of 1×103 cord blood CD34+ cells for 5 weeks with SR1 (1 μM) results in the production of 1.69×106 colony forming cells. SR1-induces CD34+ cell expansion acts by binding and antagonizing AhR as evident by decreased CYP1B1 and AHRR mRNA levels. SR1 (1 μM) treatment accelerates the proliferation of CD34+ cells and decreased the expression levels of VentX in human CD34+ cells. Ectopic expression of VentX prevents SR1-induced expansion of CD34+ cells. Sequential coculture with bone morphogenetic protein 4 (20 ng/mL), PGE2 (2 μM), and SR1 (0.75 μM) lead to robust Macaca nemestrina iPSC hematopoietic progenitor cell formation. CD34(+)CD38(-)Thy1(+)CD45RA(-)CD49f(+) cells isolated on the basis of CD34 expression and cultured in SR1 (0.75 μM) expands 3-fold and maintained this long-term repopulating HSC phenotype. StemRegenin-1(SR1)常見問題: 1. 如何使用SR1,使用濃度多少? 大多數(shù)培養(yǎng)條件下,稀釋SR1儲存液(如10 mM DMSO) 1:10,000,加入感興趣的造血干細胞培養(yǎng)基內(nèi)獲得工作液濃度為1 μM。微量濃度的DMSO(0.01%)對細胞培養(yǎng)物*。但對于非傳統(tǒng)的培養(yǎng)基,如含高濃度人血清的培養(yǎng)基,建議進行一組不同濃度SR1預(yù)實驗,因為SR1的蛋白結(jié)合特性仍未知。 2. SR1的穩(wěn)定性如何?加入培養(yǎng)基后多久需要換SR1? 細胞培養(yǎng)基內(nèi)的SR1穩(wěn)定性很好,但每2-3周需換新的SR1培養(yǎng)液。 3. 添加化合物后多久檢測SR1的作用效應(yīng)? 根據(jù)科研文獻,在7-21天內(nèi)可以觀察到SR1效應(yīng)。 4. SR1可否用于體內(nèi)實驗? 至今未有報道,但客戶可嘗試做相關(guān)實驗。 StemRegenin-1(SR1)(造血干細胞擴增劑) StemRegenin-1(SR1)的研究背景: 2010年9月Boitano等在《科學(xué)》雜志隆重介紹了其利用原代HSCs進行無偏倚篩選技術(shù)得到的一種嘌呤霉素衍生物—StemRegenin1 (SR1)。SR1是目前為止*個能促進人CD34+ 細胞大量擴增和自我更新的小分子。將SR1加入HSCs培養(yǎng)物中可以促使表達CD34的細胞數(shù)量增加50倍,而移植到免疫缺陷小鼠仍維持原來功能的HSCs細胞數(shù)增加17倍。另外,SR1還可以非常高效的促使正常HSCs或白血病干細胞/祖細胞的體外(ex vivo)擴增。因此,STR1已經(jīng)成為體外誘導(dǎo)HSC擴增的方法,也為HSCs在臨床上的應(yīng)用帶來極大方便。 華雅干細胞長期提供*的StemRegenin-1(SR1)化合物,為您的HSCs培養(yǎng)及相關(guān)研究邁出基礎(chǔ)性的一步。 |