培养基Sigma-Aldrich 干细胞培养

癌症干细胞假说

传统观点认为，癌症是环境或内部因素诱导正常细胞内的关键致癌基因和肿瘤抑制基因突变导致的一种疾病。当这些突变导致细胞向更原始、高度增殖状态转化，克隆扩张形成白血病或实体肿瘤时，就会出现癌症的临床表现¹。然而，这种模式不能够*解释许多肿瘤和转移瘤的长发育延迟，产生细胞去分化和细胞永生的起始诱导机制，或肿瘤本身内的细胞功能和表型多样性起源。过去十年中，越来越多的证据表明，恶性肿瘤起源于组织干细胞在突变作用下的转化，这些突变会导致控制干细胞生长和增殖的正常机制的调节异常^1-9。

成体干细胞填充每个器官内的离散区位。它们具有长生命期和多能性能，可以重新转化成该器官内的所有类型的细胞。干细胞的两个显著特性是：

1. 在长期的区位环境中保持休眠的能力，

2. 不对称细胞分裂能力，可分裂为一个干细胞（称作自我更新）和一个分化的子代。

支持CSC假说的例证

此外，干细胞具有细胞凋亡耐受性，具有增强的端粒酶和DNA修复活性，并且具有膜结合的ABC转运蛋白，能够排除异物，从而使干细胞相对耐受剂和其他外源性毒物的损伤作用^9-11 。自我更新加上长休眠期，使接触损伤剂的干细胞能积聚可能随着时间推移导致恶性转化的突变³。

癌症干细胞鉴定

研究人员已经从大多数白血病^2,4,13以及许多实体瘤（如脑胶质母细胞瘤和成神经管细胞瘤^4,5,10,14,15 、乳腺^3,16 、子宫颈、结肠直肠^17,18 、胃肠道、肝细胞、肺、胰腺、前列腺和皮肤癌^6,9,19,20）中分离得到少量的干细胞样细胞。癌症干细胞与其对应的正常干细胞具有相同的细胞表面标记物（见表1），但却表现出不受控制的增殖，这可能是由于对负生长调节因子或对接触抑制和间隙连接细胞间通讯（GJIC）缺失的响应性降低所致。^1,10

产品特点

AldeRed ALDH检测分析

癌症干细胞醛脱氢酶（ALDH）的表达量升高，被认为是所有CSCs的标志。ALDEFLUOR™检测试剂盒过去一直用于测量ALDH的表达水平，并广泛用于研究罕见的CSC群体。ALDEFLUOR检测试剂盒的发射波长位于电磁光谱的绿色区域（512nm）。因此，其中的试剂不能同时用于表达绿色荧光蛋白（发射绿色荧光光谱）的细胞或小鼠。AldeRed是用于标记活ALDH阳性细胞的ALDH红移荧光底物，从而释放出绿色发射通道以帮助同步完成其他检测。AldeRed克服了ALDEFLUOR检测试剂盒目前的局限。

阅读《Nature》中关于AldeRed的文章

AldeRed的特性和优点：

• 红移检测释放出其他实验所需的绿色通道

• 活体干细胞鉴定技术，能够对罕见的细胞群体进行流式分选

• 快速酶反应检测方案

AldeRed 588-A是一种无毒的ALDH荧光底物，能够自由进入完整活细胞但其被ALDH转化成酸产物后仍将滞留在细胞内。

癌症干细胞信号转导通路

干细胞巢被充满时，正常干细胞的自我更新将终止。与正常干细胞不同，癌症干细胞和干细胞源癌祖细胞的不受控自我更新会超过其区位并渗入周围组织¹³。Hedgehog，Notch，Wnt，和PTEN是控制正常和癌症干细胞自我更新、增殖和存活的部分途径。在大多数侵袭性癌症中，都可观察到导致这些途径不同程度激活的突变。

Hedgehog 和 Bmi1通路

Hedgehog（Hh）通路能够调节成体干细胞的休眠和自我更新。研究人员已经在哺乳动物中鉴定了三种Hh配体，即Sonic hedgehog（Shh）、Desert hedgehog（Dhh）和Indian hedgehog（Ihh），其中对Shh的研究多。在没有配体的情况下，Hh受体PTCH1通过催化抑制跨膜蛋白Smoothened（SMO），从而抑制信号转导。配体与PTCH1可使受体失活，并活化SMO，继而诱导Gli转录因子；Gli1和Gli2是Hh信号通路的阳性信号介导子，而Gli3是负调节子^3,20。

Bmi1的诱导能够促进许多Hh活化的效应。Bmi1 是一个的多梳基因，通过染色质重塑抑制转录，并能够下调Ink-4A / ADP核糖基化因子（ARF）复合物（如p16 Ink4A 和p19 ARF）的基因表达——这些基因是细胞周期的负调节物并且参与干细胞的休眠和分化^3,4。这使得Hh通路元件PTCH1、Gli1和Gli2表达上调^19,20， 以及Gli1和Gli2诱导的生长促进基因cyclin D1、Myc和Snail表达来实现干细胞分化和自我更新。

Notch-γ-分泌酶通路

Notch信号转导能够促进正常神经干细胞的存活和增殖，抑制它们的分化。许多癌细胞系（包括T细胞白血病，以及脑、乳腺、卵巢、宫颈、结肠直肠、胰腺、唾液腺和肺部癌症^3,6,9,15 ）的干细胞样细胞中，Notch信号转导通路被高度激活。其中，成神经管细胞瘤和T细胞淋巴细胞白血病是Notch依赖性的恶性肿瘤¹⁵。

Notch通路激活包括通过γ-分泌酶对Notch配体/受体复合物进行蛋白水解切割，从而释放Notch胞内结构域片段（NICD），移位到细胞核并上调Myc、Hes1和其他基因的表达。当用NICD2转染DAOY成神经管细胞瘤细胞系使Notch通路持续激活时，转染后的细胞比非转染的DAOY细胞产生更多的异种移植肿瘤，培养物中CD133 +和侧群干细胞样细胞也增加了。相反，抑制γ-分泌酶可将侧群细胞减少至总细胞计数的0.01％，并且抑制90％的细胞形成软琼脂集落及在免疫受损的小鼠中形成肿瘤异种移植物的能力。NICD2转染可保护细胞免受γ-分泌酶抑制的影响¹⁵。因此，在一些肿瘤类型中，抑制Notch信号转导可以消除肿瘤起始所需的细胞群。

Wnt-β-连环蛋白途径

Wnt包含结合Frizzled（Fz）受体家族的19种胞外糖蛋白家族，其可激活抑制β-连环蛋白的蛋白水解降解途径，这使得β-连环蛋白在细胞质中积累并向细胞核转运。在细胞核中，β-连环蛋白通过Tcf / Lef转录因子促进基因转录。在Wnt未激活的情况下，轴蛋白（Axin）和腺瘤 息肉病大肠杆菌蛋白（APC）与糖原合酶激酶3（Gsk3）形成复合物，从而促进细胞质β-连环蛋白磷酸化，继而靶向β-连环蛋白以进行蛋白水解降解。

Wnt通路的激活需要Fz与其共同受体（LDL受体样蛋白5/6（Lrp5 / 6））的结合。Wnt / Fz / Lrp5 / 6复合物的形成激活磷蛋白Dishevelled（Dvl）（Fz受体的组成成分），继而招募Axin / Gsk3，从而解离Axin / APC / Gsk3复合物。Lrp5 / 6被Gsk3和酪蛋白激酶1γ（Ck1γ）。 磷酸化的Lrp5 / 6结合Axin / Gsk3形成复合体，防止β-连环蛋白的磷酸化从而稳定β-连环蛋白。

图1. 癌症干细胞中关键发育信号转导通路的激活。Shh、Wnt和Notch信号转导途径与维持癌症干细胞的致瘤潜力有关。通过与Bmi1信号转导介体和其他自我更新基因（如Nanog和Sox-2）的相互作用，这些新的调节机制可以促进或抑制自我更新，从而提供癌症治疗干预的方法。

材料

Product # Image Description Molecular Formula Add to Cart A3230 AR-A014418 ≥98% (HPLC), solid C12H12N4O4S 价格 B1686 BIO ≥98% (HPLC) C16H10BrN3O2 价格 B9678 Monoclonal Anti-Bone Morphogenetic Protein 4 antibody produced in mouse clone 66119.11, purified immunoglobulin, lyophilized powder 价格 B7185 Anti-Breast Cancer Resistance Protein antibody produced in rabbit affinity isolated antibody, buffered aqueous solution 价格 B7684 Monoclonal Anti-Breast Cancer Resistance Protein antibody produced in mouse 250 μg/mL, clone BXP-34, tissue culture supernatant, aqueous solution 价格 B7059 Monoclonal Anti-Breast Cancer Resistance Protein antibody produced in mouse ~250 μg/mL, clone BXP-21, tissue culture supernatant, aqueous solution 价格 C2206 Anti-β-Catenin antibody produced in rabbit whole antiserum 价格 C7082 Monoclonal Anti-β-Catenin antibody produced in mouse clone 6F9, ascites fluid 价格 C7207 Monoclonal Anti-β-Catenin antibody produced in mouse clone 15B8, ascites fluid 价格 P7437 Monoclonal Anti-CD19−PE antibody produced in mouse clone SJ25-C1, purified immunoglobulin, buffered aqueous solution 价格 HPA005785 Anti-CD44 antibody produced in rabbit Prestige Antibodies®Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution 价格 C7923 Monoclonal Anti-CD44 antibody produced in mouse clone A3D8, purified immunoglobulin, buffered aqueous solution 价格 HPA000440 Anti-CD45 antibody produced in rabbit Prestige Antibodies®Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution 价格 C9819 Chondroitin sulfate A sodium salt from bovine trachea lyophilized powder, BioReagent, suitable for cell culture 价格 C3116 Anti-CXCR4 antibody produced in rabbit affinity isolated antibody, buffered aqueous solution 价格 C6598 Monoclonal Anti-CXCR4 (Fusin) antibody produced in mouse clone 44716, purified immunoglobulin, lyophilized powder 价格 C8352 Anti-CXCR4 (Fusin) (182-196) antibody produced in rabbit IgG fraction of antiserum, buffered aqueous solution 价格 C8227 Anti-CXCR4 (Fusin), N-Terminal antibody produced in rabbit IgG fraction of antiserum, buffered aqueous solution 价格 D5942 DAPT ≥98% (HPLC), solid C23H26F2N2O4 价格 D0817 (−)-Deguelin >98% (HPLC), powder C23H22O6 价格 E1270 ECM Gel from Engelbreth-Holm-Swarm murine sarcoma liquid, BioReagent, suitable for cell culture 价格 F3542 Fibronectin Fragment III1-C human recombinant, expressed in E. coli, lyophilized powder, ≥90% (SDS-PAGE) 价格 F3804 Anti-Frizzled-1 antibody produced in rabbit affinity isolated antibody, buffered aqueous solution 价格 F3179 Anti-Frizzled-3 antibody produced in rabbit affinity isolated antibody, buffered aqueous solution 价格 F3429 Anti-Frizzled-4 antibody produced in rabbit affinity isolated antibody, buffered aqueous solution 价格 F3679 Anti-Frizzled-7 antibody produced in rabbit affinity isolated antibody, buffered aqueous solution 价格 P7965 Monoclonal Anti-P-Glycoprotein (MDR) antibody produced in mouse clone F4, ascites fluid 价格 HPA003539 Anti-IL3RA antibody produced in rabbit Prestige Antibodies®Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution 价格 J4127 Anti-Jagged 1 antibody produced in goat affinity isolated antibody, lyophilized powder 价格 J4145 Jervine ≥98% (HPLC), powder C27H39NO3 价格 L1790 L-685,458 >96% (HPLC), solid C39H52O6N4 价格 L4020 Monoclonal Anti-LEF-1 (β-Catenin binding domain) antibody produced in mouse clone REMB1, ~1 mg/mL, purified immunoglobulin, buffered aqueous solution 价格 T2949 Anti-mTOR antibody produced in rabbit 1.0-1.5 mg/mL, affinity isolated antibody, buffered aqueous solution 价格 N8523 Monoclonal Anti-NF-κB antibody produced in mouse clone NF-12, ascites fluid 价格 N6786 Monoclonal Anti-Notch1 antibody produced in mouse clone mN1A, purified immunoglobulin, buffered aqueous solution 价格 N9286 Notch-1/Fc Chimera from rat recombinant, expressed in NSO cells, ≥90% (SDS-PAGE), lyophilized powder, from rat 价格 N9161 Notch-2/Fc Chimera from rat recombinant, expressed in NSO cells, ≥90% (SDS-PAGE), lyophilized powder 价格 P4235 Anti-p53 antibody produced in sheep fractionated antiserum, buffered aqueous solution 价格 P5813 Monoclonal Anti-p53 antibody produced in mouse clone BP53-12, ascites fluid 价格 P8999 Monoclonal Anti-p53 antibody produced in mouse clone DO-7, purified from hybridoma cell culture 价格 P7874 Monoclonal Anti-p53 antibody produced in mouse ~2 mg/mL, clone DO-2, purified immunoglobulin, buffered aqueous solution 价格 P6874 Monoclonal Anti-p53 antibody produced in mouse clone DO-1, purified from hybridoma cell culture 价格 P0667 Parthenolide ≥98% (HPLC) C15H20O3 价格 P4707 Poly-L-lysine solution mol wt 70,000-150,000, 0.01%, sterile-filtered, BioReagent, suitable for cell culture 价格 P4832 Poly-L-lysine solution mol wt 150,000-300,000, 0.01%, sterile-filtered, BioReagent, suitable for cell culture 价格 P7482 Anti-PTEN antibody produced in rabbit IgG fraction of antiserum, buffered aqueous solution 价格 P3487 Monoclonal Anti-PTEN antibody produced in mouse ~1 mg/mL, purified immunoglobulin, clone PTN-18, buffered aqueous solution 价格 R1276 Reversin 121 ≥97% (HPLC), powder C34H47N3O9 价格 R1401 Reversin 205 ≥98% (HPLC), film C41H58N4O12 价格 S4572 SANT-1 ≥98% (HPLC), powder C23H27N5 价格 S2188 γ-Secretase inhibitor ≥90% (TLC), powder C33H57F2N5O9 价格 S9819 Anti-Smoothened Drosophila Homolog antibody produced in rabbit affinity isolated antibody 价格 S0191 Sonic Hedgehog Peptide from mouse ≥97% (SDS-PAGE), recombinant, expressed in E. coli, Fused to a histidine tag at the C-terminus, lyophilized powder 价格 S8321 Monoclonal Anti-Sonic Hedgehog (Shh) 小鼠抗 clone SH154, purified from hybridoma cell culture 价格 S4944 Monoclonal Anti-Sonic Hedgehog (N-terminal) antibody produced in rat clone 171018, purified immunoglobulin, lyophilized powder 价格 S5174 SPARC from murine parietal yolk sac (PYS-2) cells ~80% (SDS-PAGE), lyophilized powder 价格 HPA001671 Anti-STAT3 antibody produced in rabbit Prestige Antibodies®Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution 价格 S5933 Anti-STAT3 antibody produced in rabbit IgG fraction of antiserum, buffered aqueous solution 价格 V1014 Anti-Vascular Endothelial Growth Factor Receptor 2 antibody produced in goat affinity isolated antibody, lyophilized powder 价格 V8379 Vitronectin 来源于人类血浆 lyophilized powder, BioReagent, suitable for cell culture 价格 W2266 Anti-Wnt-4 antibody produced in goat affinity isolated antibody, lyophilized powder 价格 P7280 多聚-D-赖氨酸 氢溴酸盐 average mol wt 30,000-70,000, lyophilized powder, γ-irradiated, BioReagent, suitable for cell culture 价格 P6407 多聚-D-赖氨酸 氢溴酸盐 mol wt 70,000-150,000, lyophilized powder, γ-irradiated, BioReagent, suitable for cell culture 价格 L2020 层粘蛋白 来源于 Engelbreth-Holm-Swarm 小鼠肉瘤基底膜 1-2 mg/mL in Tris buffered NaCl, 0.2 μm filtered, BioReagent, suitable for cell culture 价格 L6274 层粘蛋白 来源于人类胎盘 liquid, BioReagent, suitable for cell culture 价格 C4116 环巴胺 水合物 ≥98% (HPLC) C27H41NO2 · xH2O 价格 F2006 Fibronectin human plasma 纤连蛋白 溶液 来源于人类成纤维细胞 lyophilized powder, BioReagent, suitable for cell culture 价格 F0895 Fibronectin human plasma 纤连蛋白 溶液 来源于人类成纤维细胞 liquid, 0.1% (Solution), BioReagent, suitable for cell culture 价格 F0635 Fibronectin rat plasma 纤连蛋白 溶液 来源于人类成纤维细胞 powder, BioReagent, suitable for cell culture 价格 P9155 聚-L-赖氨酸 氢溴酸盐 mol wt 30,000-70,000, lyophilized powder, γ-irradiated, BioXtra, suitable for cell culture 价格 P6282 聚-L-赖氨酸 氢溴酸盐 mol wt 70,000-150,000, lyophilized powder, γ-irradiated, BioXtra, suitable for cell culture 价格 P5899 聚-L-赖氨酸 氢溴酸盐 mol wt ≥300,000, lyophilized powder, γ-irradiated, BioXtra, suitable for cell culture 价格 P4957 聚-L-鸟氨酸 溶液 mol wt 30,000-70,000, 0.01%, sterile-filtered, BioReagent, suitable for cell culture 价格 C5533 胶原 来源于人类胎盘 Bornstein and Traub Type IV, powder, BioReagent, suitable for cell culture 价格 C7661 胶原 来源于大鼠尾 Bornstein and Traub Type I, powder, BioReagent, suitable for cell culture 价格

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