干细胞表面标记
植物干细胞分选方法
植物干细胞分选方法植物干细胞是一种具有自我更新和分化潜能的细胞，能够在植物生长过程中不断地产生新细胞并形成不同器官和组织。
植物干细胞对植物生长和发育起着至关重要的作用，因此对植物干细胞的研究受到了广泛关注。
植物干细胞分选方法是研究植物干细胞的重要手段之一，能够帮助科研人员快速准确地获取纯净的植物干细胞，为植物干细胞的研究提供了基础条件。
植物干细胞分选方法的发展历程植物干细胞分选方法的发展经历了多个阶段。
早期的植物干细胞分选方法主要是依靠显微镜手工分选，这种方法操作繁琐、效率低下，并且容易受到环境和操作者技术水平的影响。
随着生物技术和细胞分离技术的不断发展，研究人员陆续提出了多种新的植物干细胞分选方法，包括细胞表面标记、细胞自动分选和细胞排序等技术。
这些新兴的分选方法大大提高了植物干细胞的纯度和分选效率，为植物干细胞的研究提供了更为可靠的技术支持。
植物干细胞分选方法的基本原理植物干细胞分选方法的基本原理是利用植物干细胞在生长和发育过程中的特定特征，通过物理、化学或生物学手段将植物干细胞与其他细胞进行有效分离和纯化。
常见的分选方法包括细胞表面标记法、荧光激活细胞分选法、细胞自动分选技术等。
细胞表面标记法是应用最为广泛的一种方法，其原理是利用特异性抗体或荧光标记物将植物干细胞特异性地标记出来，然后通过细胞分选设备将标记的植物干细胞与其他细胞进行分离。
这种方法操作简单、高效，能够实现快速准确地分选植物干细胞。
植物干细胞分选方法的关键技术植物干细胞分选方法的关键技术主要包括标记试剂的选择、标记条件的优化、细胞分选设备的使用等。
在植物干细胞分选过程中，选择合适的标记试剂对于分选效果起着至关重要的作用。
通常情况下，标记试剂需要具有与植物干细胞表面特异性结合的能力，并且不会对植物干细胞的生理功能产生不利影响。
优化标记条件也是确保分选效果的重要因素，如标记时间、标记温度、标记浓度等都会对植物干细胞的分选结果产生影响。
人间充质干细胞表面标记分子的研究进展
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干细胞表面标准(一)
干细胞表面标准(一)干细胞表面标准引言干细胞具有自我更新和多向分化的能力，被广泛应用于生物医学领域。
然而，由于干细胞存在不确定性和低纯度等问题，研究人员们急需一套干细胞表面标准来准确鉴定和纯化干细胞群体。
重要性干细胞表面标准对于干细胞研究和应用具有重要意义： - 纯化：标准可以帮助研究人员清晰地区分干细胞和非干细胞群体，从而纯化干细胞样品。
- 鉴定：标准可以用于鉴定干细胞的特殊表面特征，确保所研究或使用的细胞具有干细胞的属性。
- 质量控制：标准可以帮助研究人员评估干细胞样品的纯度和质量，从而确保实验结果的准确性和可重复性。
国际标准目前，国际上已被广泛接受的干细胞表面标准主要包括以下几种：- CD标记物：CD34、CD133等 - SSEA标记物：SSEA-1、SSEA-3、SSEA-4等 - 表面受体标记：CD44、CD105、CD29等标准的选择在实际应用中，选择合适的干细胞表面标准需要考虑以下几个因素： 1. 干细胞类型：不同类型的干细胞具有不同的表面标志物，选择适合特定干细胞类型的标准非常重要。
2. 研究目的：标准的选择也要根据研究目的进行，不同标准可能适用于不同的实验设计和应用需求。
3. 实验条件：在实验条件下，一些标准可能更容易使用和检测，而另一些标准可能受到技术限制。
标准的验证选择合适的干细胞表面标准后，需要进行验证确保其准确性和可靠性： 1. 流式细胞术：通过流式细胞仪可以分析细胞表面标志物的表达情况，确保标准的有效性。
2. 免疫组化：利用免疫组化技术可以观察细胞表面标志物的分布情况，验证标准的准确性。
3. 功能性鉴定：通过诱导干细胞分化为特定细胞类型，确定细胞表面标准的功能和特异性。
结论干细胞表面标准在干细胞研究和应用中起着至关重要的作用。
选择合适的标准、进行有效的验证以及在实验中严格遵循标准操作程序，将有助于确保干细胞的纯度和质量，推动干细胞领域的发展和应用。
干细胞培养中的细胞鉴定与鉴定方法
干细胞培养中的细胞鉴定与鉴定方法细胞鉴定是干细胞培养中至关重要的一步，它能够帮助科研人员确定细胞的纯度和特性，从而保证实验结果的准确性和可靠性。
在干细胞研究中，细胞鉴定的主要目的是检测细胞的多能性和细胞表面标记物的表达情况。
本文将介绍干细胞培养中常用的细胞鉴定方法，以及在干细胞培养中细胞鉴定的重要性。
一、细胞鉴定的重要性在干细胞培养中，细胞鉴定是确保所使用的细胞符合实验要求的关键步骤。
干细胞的特性和功能对于各种研究和应用领域具有重要意义，如再生医学、药物筛选、疾病模型的构建等。
因此，细胞鉴定的准确性对于确定细胞的纯度和特性是至关重要的。
细胞鉴定能够帮助研究人员确认所使用的细胞群体中是否存在其他非目标细胞类型的杂质。
通过检测细胞上的特定标记物，可以确定细胞的起源和特性，从而准确判断细胞的多能性。
此外，细胞鉴定还可以帮助研究人员监测细胞培养过程中的质量控制，及时发现并排除异常细胞。
细胞鉴定的结果直接影响着后续实验的设计和结果解释。
准确地进行细胞鉴定可以确保实验数据的可靠性和可重复性，而不准确的细胞鉴定可能会导致误导性的结论或不可靠的实验结果。
因此，在干细胞培养中，细胞鉴定是不可或缺的步骤，它有助于保证实验结果的准确性和可靠性。
二、常用的细胞鉴定方法在干细胞培养中，常用的细胞鉴定方法包括细胞表面标记物的检测和多能性的评估。
1.细胞表面标记物的检测通过检测细胞表面标记物的表达情况，可以确定细胞类型和特性。
常用的细胞鉴定方法包括流式细胞术（flow cytometry）、免疫细胞化学（immunocytochemistry）和细胞表面蛋白质鉴定（surface protein identification）。
流式细胞术常用于检测细胞表面蛋白质标记物的表达情况。
通过特定的抗体与细胞表面标记物结合，并使用荧光染料标记抗体来实现对标记物的定量检测。
该方法可以同时检测多个标记物，高通量检测，准确鉴定细胞群体中的不同细胞类型。
造血干细胞表面标志物
造血干细胞表面标志物造血干细胞(Hematopoietic stem cells, HSC)是一类成体干细胞，具有干细胞的特性：自我更新和分化潜能。
造血干细胞是一种骨髓来源的多能干细胞，它是血液系统中的“种子”细胞。
造血干细胞(HSCs)在造血过程中形成血系中的所有细胞，包括各种成熟细胞如白细胞、红细胞、血小板等。
在受到适当刺激时还分化为其他非造血组织(如脂肪细胞、心肌细胞、内皮细胞和胰腺细胞)[1]。
造血干细胞具有调节体内平衡、免疫功能、抗微生物、抗炎症等生物学功能。
它在血液病、遗传性血液病和自身免疫性疾病的治疗方面有重要作用。
1. 造血干细胞类型2. 造血干细胞的来源3. 造血干细胞的生物学特性4. HSC和免疫系统5. 人体造血系统层次6. 造血干细胞的细胞表面标志7. HSC的临床应用1. 造血干细胞类型已经定义了两种类型的造血干细胞：能够终生保持自我更新和多谱系分化潜能的长期再生细胞(LTRC)；来源于LTRC的短期再生细胞(STRC)，虽然它们保持了多能性，但它们表现出更有限的自我更新潜能。
它们重建髓系和/或淋巴系间隔的时间很短，大约6周。
2. 造血干细胞的来源造血干细胞存在于成年人的骨髓中，特别是在骨盆、股骨和胸骨中。
它们也存在于脐带血和少量的外周血中。
2.1 骨髓造血干细胞(HSCs)是一种骨髓来源的多能干细胞。
从骨髓中获取造血干细胞通过外科手术，从两个髂骨后嵴分次采集。
骨髓中每10万个细胞中约有1个是长期造血干细胞(LT-HSC)。
2.2 外周血大部分造血干细胞来源于骨髓，少量的干细胞和祖细胞在血液中循环。
人类造血干细胞的临床移植，可以从外周血中收集供体细胞。
造血干细胞的采集是在粒细胞集落刺激因子(G-CSF)等造血生长因子的作用下，将骨髓中的HSC动员到外周血后，通过分离的方式采集。
2.3 脐带血(UCB)脐带血是造血干细胞和造血祖细胞的丰富来源，它所含不同类型的造血祖细胞的数量大约是成人血液中观察到的数量的10倍。
小鼠干细胞marker基因-概述说明以及解释
小鼠干细胞marker基因-概述说明以及解释1.引言1.1 概述小鼠干细胞是一类特殊的细胞，具有自我更新能力和多向分化潜能。
它们在发育过程中起着关键作用，并且被广泛应用于生物医学研究领域。
为了更好地理解和利用小鼠干细胞，科学家们致力于寻找能够准确标记这些干细胞的特殊基因，即干细胞marker基因。
干细胞marker基因是指在干细胞中高表达的基因，其特异性表达可以用来鉴定和分离干细胞群体。
这些基因的研究对于小鼠干细胞的定位、纯化以及研究其分子机制等方面起到至关重要的作用。
通过研究已知的小鼠干细胞marker基因，我们可以更好地了解小鼠干细胞的特征和功能。
已知的marker基因包括Oct4、Sox2、Nanog等，它们在小鼠干细胞中表达较高，而在非干细胞中表达较低或不表达。
这些基因的发现，为我们准确鉴定和纯化小鼠干细胞提供了重要依据，并促进了对小鼠干细胞自我更新和分化调控机制的进一步研究。
尽管已知的小鼠干细胞marker基因已经有了一定的认识，但仍然有许多未知的marker基因等待我们去探索。
未来的研究方向是通过新的技术手段和策略，发现更多具有特异性表达的marker基因，从而全面揭示小鼠干细胞的特性和功能。
综上所述，小鼠干细胞marker基因研究具有重要的意义。
它为我们深入了解小鼠干细胞提供了基础，并为干细胞相关的疾病治疗和组织工程等领域的研究提供了指导和突破口。
未来的研究将进一步延伸我们对小鼠干细胞的认识，促进干细胞领域的科学发展和应用。
1.2文章结构文章结构部分的内容可以按照以下方式编写：文章结构：本篇文章分为引言、正文和结论三个部分。
引言部分将对小鼠干细胞marker基因进行概述，并介绍本文的文章结构和目的。
正文部分分为三个部分：小鼠干细胞的定义和特点、干细胞marker 基因的重要性以及已知的小鼠干细胞marker基因。
在小鼠干细胞的定义和特点部分，将介绍小鼠干细胞的来源、特点以及其在生物学研究中的重要性。
干细胞标记物
受体：（receptor）是一种能够识别和选择 性结合某种配体（信号分子）的大 分子物质
受体与配体间的作用具有三个主要特征： ①特异性； ②饱和性； ③高度的亲和力。
白细胞分化抗原： 是白细胞（还包括血小板、血管内皮细胞
等）在分化成熟为不同谱系（lin-eage）和分 化不同阶段以及活化过程中，出现或消失的细 胞表面标记
当造血干细胞分化为各系的祖细胞和成熟细胞时，会出现各系细胞的专一标 志，即系表面抗原 如： 淋巴 系：CD19、CD7 粒 系：CD11、CD13、CD15、CD16 红 系：CD36、CD47、CD51、CD59、CD71； 单核 系：CD14 巨核 系：CD31、CD41、CD42、CD61、CD107 B淋巴系：CD19、CD20、CD21、CD22 T/NK 系：CD2、CD7、CD11、CD25、CD26
粘附分子: 是一类能介导细胞间及细胞与细胞外
基质粘附的分子，多为糖蛋白。
粘附分子分类: 粘合素超家族粘附分子（纤维粘连蛋白、层粘连 蛋白、纤维蛋白原、胶原蛋白、等） 免疫求蛋白超家族粘附分子（ICAM、VCAM、和CD31等） 钙粘连素超家族粘附分子 选择素家族粘附分子 其它粘附分子。
干细胞表面分子功能： 参与干细胞的分化调控 维持干细胞的自我更新和多能性 参与机体重要的生理和病理过程
Kit：原癌基因c-kit表达产物（又叫干细胞因子受 体SCF-R,CD117）,表达于几乎
所有小鼠造血干细胞。 C-kit可编码一种穿膜酪氨酸激酶受体分子。 存在于造血干细胞膜上，它的配体分子是造血干细胞因子（stem cell
factor,SCF）。 人类60%-75%的CD34+造血细胞同时表达c-kit分子。kit+细胞占骨髓细胞的1%-4%，
干细胞标记物
组织特异性标记物
EpCAM
上皮细胞黏附分子，表达 于上皮来源的干细胞和祖 细胞，如乳腺干细胞、肝 干细胞等。
Nestin
一种中间丝蛋白，表达于 神经干细胞、肌肉卫星细 胞等，参与细胞骨架构成 和细胞迁移。
Sox2
一种转录因子，表达于神 经干细胞和祖细胞，参与 神经系统的发育和维持。
功能性标记物
1 2
干细胞标记物
汇报人：XX
目录
• 干细胞标记物概述 • 干细胞表面标记物 • 干细胞内部标记物 • 干细胞标记物检测技术与方法 • 干细胞标记物在再生医学中的应
用 • 干细胞标记物在疾病诊断与治疗
中的应用
01
干细胞标记物概述
定义与分类
干细胞标记物定义
干细胞标记物是指在干细胞中特异性 表达或高表达的蛋白质、基因或糖类 等物质，用于干细胞的鉴定、分离和 纯化。
超声成像技术
利用超声成像技术对干细胞进行实时监测，观察干细胞的增殖、 分化和组织再生情况。
05
干细胞标记物在再生医学 中的应用
组织工程领域应用
种子细胞来源
干细胞作为组织工程中的种子细胞，可通过标记物鉴定其类型、纯 度和分化潜能。
细胞增殖与分化调控
利用干细胞标记物可监测干细胞的增殖和分化过程，为组织工程提 供最佳的细胞生长条件。
移பைடு நூலகம்后功能恢复
干细胞标记物可用于监测移植后器官的功能恢复 情况，为移植患者提供个性化的治疗方案。
细胞治疗领域应用
细胞替代治疗
利用干细胞标记物可鉴定出具有特定功能的干细胞，用于 替代受损或病变的细胞，恢复组织或器官的正常功能。
基因治疗载体
干细胞作为基因治疗的理想载体，可通过标记物鉴定其转 染效率和基因表达情况，为基因治疗提供可靠的细胞来源 。
BD人间充质干细胞表面标记检测试剂盒说明书
BD Stemflow™Technical Data SheetHuman MSC Analysis KitProduct InformationMaterial Number:562245Size: 50 testsConfirmed: HumanReactivity:DescriptionHuman multipotent mesenchymal stromal cells (MSCs), also referred to as mesenchymal stem cells, are a rare population of adult stem cellsthat can be isolated from a variety of tissues. MSCs that have been isolated from bone marrow and subsequently cultured can differentiate to avariety of cell types, most notably adipocytes, osteocytes and chondrocytes. MSCs also have immunomodulatory effects in vivo and in vitro.In 2006, the International Society for Cellular Therapy (ISCT) proposed a cell surface marker panel for the minimal identification of humanMSCs derived from bone marrow. Under this recommendation MSCs should be positive for CD73, CD90, and CD105, but be negative forCD34, CD45, CD11b or CD14, CD19 or CD79α, and HLA-DR. MSCs are also known to express numerous cell surface markers such asCD44, CD29, CD200, CD166, CD146 and CD271. In this kit we include the panel that was proposed by the ISCT. In addition, we designedthis kit to be modular so that additional markers could be “dropped-in”. Specifically, the MSC positive cocktail (FITC CD90, PerCP-Cy™5.5CD105 and APC CD73) leaves the PE channel open to use in combination with the supplied negative MSC cocktail (PE CD45, PE CD34, PECD11b, PE CD19 and PE HLA-DR), the included PE CD44 antibody conjugate, or a variety of other commercially available PE antibodyconjugates. Multi color analysis minimizes cell numbers required for an assay and antibody cocktails facilitate a streamlined staining protocolto analyze multiple samples. Individual positive markers are included for compensation set-up.Kit ComponentsComponent Description Size Vol. Per Test Storage Buffer51-9007661PE hMSC Negative Cocktail50 Test20 µl Aqueous buffered solution containingBSA and ≤0.09% sodium azide51-9007662PE hMSC Isotype Control Negative Cocktail50 Test20 µl Aqueous buffered solution containingBSA and ≤0.09% sodium azide51-9007663hMSC Positive Cocktail50 Test20 µl Aqueous buffered solution containingBSA and ≤0.09% sodium azide51-9007664hMSC Isotype Control Positive Cocktail50 Test20 µl Aqueous buffered solution containingBSA and ≤0.09% sodium azide51-9007657FITC Mouse Anti-Human CD9050 Test 5 µl Aqueous buffered solution containingBSA and ≤0.09% sodium azide51-9007656PE Mouse Anti-Human CD44100 Test 5 µl Aqueous buffered solution containingBSA and ≤0.09% sodium azide51-9007655PE Mouse IgG2b, k Isotype Control50 Test 5 µl Aqueous buffered solution containingBSA and ≤0.09% sodium azide51-9007649APC Mouse Anti-Human CD7350 Test 5 µl Aqueous buffered solution containingBSA and ≤0.09% sodium azide51-9007648PerCP-Cy™5.5 Mouse Anti-Human CD10550 Test 5 µl Aqueous buffered solution containing(Endoglin)BSA and ≤0.09% sodium azideDescription of Kit ComponentsVial Contents PurposehMSC Positive Cocktail CD90 FITC (Clone: 5E10)Cocktail to positively identify hMSCsCD105 PerCP-Cy5.5 (Clone: 266)CD73 APC (Clone: AD2)hMSC Positive Isotype Control Cocktail mIgG1, κ FITC (Clone: X40)Corresponding Isotype Control for hMSC PositivemIgG1, κ PerCP-Cy5.5 (Clone: X40)CocktailmIgG1, κ APC (Clone: X40)PE hMSC Negative Cocktail CD34 PE (Clone:581)Cocktail to identify potential contaminantsCD11b PE (Clone: ICRF44)CD19 PE (Clone: HIB19)CD45 PE (Clone: HI30)HLA-DR PE (Clone: G46-6)PE hMSC Negative Isotype Control Cocktail mIgG1, κ PE (Clone: X40)Corresponding isotype control for PE hMSCmIgG2a, κ PE (Clone:G155-178)Negative CocktailFITC Mouse Anti-human CD90CD90 FITC (Clone: 5E10)Compensation controlPE Mouse Anti-Human CD44CD44 PE (Clone: G44-26)Compensation control/MSC positive drop-in PerCP-Cy™5.5 Mouse Anti-Human CD105CD105 PerCP-Cy™5.5 (Clone: 266)Compensation controlAPC Mouse Anti-Human CD73CD73 APC (Clone:AD2)Compensation controlPE Mouse IgG2b, κ Isotype Control mIgG2b, κ (Clone: 27-35)Corresponding Isotype Control for PE MouseAnti-Human CD44, when used as a drop in Preparation and StorageStore undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.The antibody was conjugated with FITC under optimum conditions, and unreacted FITC was removed.The antibody was conjugated to APC under optimum conditions, and unconjugated antibody and free APC were removed.Application NotesApplicationFlow cytometry Tested During DevelopmentRecommended Assay Procedure:(1) Detach cells using BD™ Accutase™ Cell Detachment Solution (Cat. No 561527) or similar detachment solution, wash cells and resuspend ata concentration of 1x10^7 cells/ml in BD Pharmingen™ Stain Buffer (Cat. No. 554656) or other appropriate staining buffer.(a) Cells can also be resuspended at a concentration of 5x10^6 cells/ml if cell number is a limiting factor.(2) Label tubes and add antibodies as shown below:Tube Add (1 test size)(1)FITC Mouse Anti-Human CD90 (5µl)(2)PE Mouse Anti-Human CD44 (5µl)(3)PerCP-Cy™5.5 Mouse Anti-Human CD105 (5µl)(4)APC Mouse Anti-Human CD73 (5µl)(5)Nothing(6)hMSC Positive Isotype Control Cocktail (20µl)PE hMSC Negative Isotype Control Cocktail (20µl)(7)hMSC Positive Cocktail (20µl)PE hMSC Negative Cocktail (20µl)And/or(8)hMSC Positive Isotype Control Cocktail (20µl)Drop in isotype control (i.e. PE Mouse IgG2b, κ) (5 µl)(9)hMSC Positive Cocktail (20µl)PE Drop in (i.e. PE Mouse Anti-Human CD44) (5µl)(3) Repeat tubes 5-7 and/or 8-9 for each additional cell sample.(4) Add 100 µl of prepared cell suspension to tubes 1 through 9.(5) Incubate tubes in the dark for 30 minutes (May be done on ice or at room temp)(6) Wash the cells twice with BD Pharmingen™ Stain Buffer (FBS) and resuspend at 300-500 µl in BD Pharmingen™ Stain Buffer (FBS) . Alternatively, 1X Washing/Staining Solution (1 x PBS,1% FCS, and 0.09% sodium azide) may be used.(7) Analyze cells on flow cytometer(a) tubes 1-5 are to be used as controls to set up cytometer (i.e Compensation)Suggested Companion ProductsSizeCatalog Number Name Clone 554656Stain Buffer (FBS)500 ml(none)(none)561527Accutase™ Cell Detachment Solution100 mlTop: Schematic workflow of the human MSC analysis kit.Bottom: Cells grown in BD Mosaic™ hMSC SF Cell Culture Environment (Cat. No. 355700) were detached using BD™ Accutase™ CellDetachment Solution (Cat. No 561527) and then stained with the positive and negative cocktails (solid lines) or the positive and negativeisotype control cocktails (dashed lines). The plots were derived from gated events based on light scattering characteristics of the MSCs.Cells were analyzed using a BD™ LSRII flow cytometry system.Product Notices1.Accutase is a registered trademark of Innovative Cell Technologies, Inc.2.Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.3.This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the4.tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant5.spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos.6.5,486,616; 5,569,587; 5,569,766; 5,627,027.This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under7.license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.8.Source of all serum proteins is from USDA inspected abattoirs located in the United States.9.Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water beforediscarding to avoid accumulation of potentially explosive deposits in plumbing.10.For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at/colors.11.Please refer to /pharmingen/protocols for technical protocols.ReferencesBühring HJ, Battula VL, Treml S, Schewe B, Kanz L, Vogel W. Novel markers for the prospective isolation of human MSC. Ann N Y Acad Sci. 2007;1106:262-271. (Biology: Flow cytometry)Dominici M, Le Blanc K, Mueller I, et. al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006; 8(4):315-317. (Biology: Flow cytometry)Horwitz EM, Le Blanc K, Dominici M, et. al. Clarification of the nomenclature for MSC: The International Society for Cellular Therapy position statement.Cytotherapy. 2005; 7(5):393-395. (Biology)Kern S, Eichler H, Stoeve J, Klüter H, Bieback K. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue.Stem Cells. 2006; 24(5):1294-1301. (Biology: Flow cytometry)Sánchez L, Gutierrez-Aranda I, Ligero G, et al. Enrichment of Human ESC-Derived Multipotent Mesenchymal Stem Cells with Immunosuppressive andAnti-Inflammatory Properties Capable to Protect Against Experimental Inflammatory Bowel Disease. Stem Cells. 2010; 29(2):251-262. (Biology: Cell differentiation)。
免疫荧光法检测干细胞标志物
免疫荧光法检测干细胞标志物英文回答：Immunofluorescence is a widely used technique in the field of cell biology to detect and visualize specific proteins or molecules of interest within cells. This technique involves the use of antibodies that are labeled with fluorescent dyes, which emit light of specific wavelengths when excited by a specific light source. By targeting specific cell surface markers or intracellular proteins, immunofluorescence allows us to identify and characterize different cell types or specific cellular components.To detect stem cell markers using immunofluorescence, we first need to identify the specific markers that are expressed on the surface of stem cells. For example, in the case of embryonic stem cells, the markers Oct4, Nanog, and Sox2 are commonly used. Once we have identified the markers of interest, we can proceed with the immunofluorescenceprotocol.The first step is to fix the cells using a fixativesuch as paraformaldehyde, which helps to preserve thecellular structure and maintain the integrity of the proteins. After fixation, the cells are permeabilized using a detergent to allow the antibodies to penetrate the cell membrane and access the intracellular proteins.Next, we incubate the cells with primary antibodiesthat specifically recognize the stem cell markers of interest. These primary antibodies are typically raised in animals such as mice or rabbits and are designed to bind specifically to the target proteins. After incubation with the primary antibodies, we wash away any unbound antibodies.The next step is to add secondary antibodies that are conjugated with fluorescent dyes. These secondaryantibodies recognize and bind to the primary antibodies, amplifying the fluorescent signal. Different fluorescent dyes can be used to label different primary antibodies, allowing for the simultaneous detection of multiple markersin the same sample.Finally, the cells are counterstained with a nuclear stain, such as DAPI, to visualize the cell nuclei. This helps to identify individual cells and provides a reference for the localization of the stem cell markers within the cells.The samples are then examined under a fluorescence microscope, and the emitted fluorescence is captured using specific filters. The resulting images can be analyzed and quantified using image analysis software to determine the expression levels and localization patterns of the stemcell markers.Immunofluorescence is a powerful technique that allows for the visualization and characterization of stem cell markers. It provides valuable information about thepresence and distribution of specific proteins within stem cells, helping researchers to better understand theirbiology and potential applications in regenerative medicine.中文回答：免疫荧光法是细胞生物学领域中广泛使用的一种技术，用于检测和可视化细胞内特定的蛋白质或分子。
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Blood VesselFetal liver kinase-1 (Flk1) Endothelial Cell-surface receptor protein that identifies endothelial cell progenitor;marker of cell-cell contactsSmooth muscle cell-specificmyosin heavy chainSmooth muscle Identifies smooth muscle cells in the wall of blood vesselsVascular endothelial cellcadherinSmooth muscle Identifies smooth muscle cells in the wall of blood vesselsBoneBone-specific alkaline phosphatase (BAP) Osteoblast Enzymeexpressed in osteoblast; activity indicates bone formationHydroxyapatite Osteoblast Minerlized bone matrix that provides structural integrity; marker ofbone formationOsteocalcin (OC) Osteoblast Mineral-binding protein uniquely synthesized by osteoblast; marker ofbone formationBone Marrow and BloodBone morphogenetic protein receptor (BMPR) Mesenchymal stem andprogenitor cellsImportant for the differentiation of committed mesenchymal cell typesfrom mesenchymal stem and progenitor cells; BMPR identifies earlymesenchymal lineages (stem and progenitor cells)CD4 and CD8 White blood cell (WBC) Cell-surface protein markers specific for mature T lymphocyte (WBCsubtype)CD34 Hematopoietic stem cell(HSC), satellite,endothelial progenitor Cell-surface protein on bone marrow cell, indicative of a HSC and endothelial progenitor; CD34 also identifies muscle satellite, a muscle stem cellCD34+Sca1+ Lin- profile Mesencyhmal stem cell(MSC) Identifies MSCs, which can differentiate into adipocyte, osteocyte, chondrocyte, and myocyteCD38 Absent on HSCPresent on WBC lineages Cell-surface molecule that identifies WBC lineages. Selection of CD34+/CD38- cells allows for purification of HSC populationsCD44 Mesenchymal A type of cell-adhesion molecule used to identify specific types ofmesenchymal cellsc-Kit HSC, MSC Cell-surface receptor on BM cell types that identifies HSC and MSC;binding by fetal calf serum (FCS) enhances proliferation of ES cells,HSCs, MSCs, and hematopoietic progenitor cellsColony-forming unit (CFU) HSC, MSC progenitor CFU assay detects the ability of a single stem cell or progenitor cell togive rise to one or more cell lineages, such as red blood cell (RBC)and/or white blood cell (WBC) lineagesFibroblast colony-forming unit (CFU-F) Bone marrow fibroblast An individual bone marrow cell that has given rise to a colony ofmultipotent fibroblastic cells; such identified cells are precursors ofdifferentiated mesenchymal lineagesHoechst dye Absent on HSC Fluorescent dye that binds DNA; HSC extrudes the dye and stainslightly compared with other cell typesLeukocyte common antigen(CD45)WBC Cell-surface protein on WBC progenitorLineage surface antigen(Lin)HSC, MSCDifferentiated RBC andWBC lineages Thirteen to 14 different cell-surface proteins that are markers of mature blood cell lineages; detection of Lin-negative cells assists in the purification of HSC and hematopoietic progenitor populationsMac-1 WBC Cell-surface protein specific for mature granulocyte and macrophage(WBC subtypes)Muc-18 (CD146) Bone marrow fibroblasts,endothelial Cell-surface protein (immunoglobulin superfamily) found on bone marrow fibroblasts, which may be important in hematopoiesis; a subpopulation of Muc-18+ cells are mesenchymal precursorsStem cell antigen (Sca-1) HSC, MSC Cell-surface protein on bone marrow (BM) cell, indicative of HSC andMSC Bone Marrow and Blood cont.Stro-1 antigen Stromal (mesenchymal)precursor cells,hematopoietic cells Cell-surface glycoprotein on subsets of bone marrow stromal (mesenchymal) cells; selection of Stro-1+ cells assists in isolating mesenchymal precursor cells, which are multipotent cells that give rise to adipocytes, osteocytes, smooth myocytes, fibroblasts, chondrocytes, and blood cellsThy-1 HSC, MSC Cell-surface protein; negative or low detection is suggestive of HSCCartilageCollagen types II and IV Chondrocyte Structural proteins produced specifically by chondrocyteKeratin Keratinocyte Principal protein of skin; identifies differentiated keratinocyte Sulfated proteoglycan Chondrocyte Molecule found in connective tissues; synthesized by chondrocyteFatAdipocyte lipid-bindingprotein (ALBP)Adipocyte Lipid-binding protein located specifically in adipocyteFatty acid transporter (FAT) Adipocyte Transport molecule located specifically in adipocyteAdipocyte lipid-bindingprotein (ALBP)Adipocyte Lipid-binding protein located specifically in adipocyteGeneralY chromosome Male cells Male-specific chromosome used in labeling and detecting donor cells infemale transplant recipientsKaryotype Most cell types Analysis of chromosome structure and number in a cellLiverAlbumin Hepatocyte Principal protein produced by the liver; indicates functioning ofmaturing and fully differentiated hepatocytesB-1 integrin Hepatocyte Cell-adhesion molecule important in cell-cell interactions; markerexpressed during development of liverNervous SystemCD133 Neural stem cell, HSC Cell-surface protein that identifies neural stem cells, which give rise toneurons and glial cellsGlial fibrillary acidic protein(GFAP)Astrocyte Protein specifically produced by astrocyteMicrotubule-associated protein-2 (MAP-2) Neuron Dendrite-specific MAP; protein found specifically in dendritic branchingof neuronMyelin basic protein (MPB) Oligodendrocyte Protein produced by mature oligodendrocytes; located in the myelinsheath surrounding neuronal structuresNestin NeuralprogenitorIntermediatefilamentstructural protein expressed in primitive neuraltissueNeural tubulin Neuron Important structural protein for neuron; identifies differentiatedneuronNeurofilament (NF) Neuron Important structural protein for neuron; identifies differentiatedneuronNeurosphere Embryoid body (EB), ES Cluster of primitive neural cells in culture of differentiating ES cells;indicates presence of early neurons and gliaNoggin Neuron A neuron-specific gene expressed during the development of neurons O4 Oligodendrocyte Cell-surface marker on immature, developing oligodendrocyteO1 Oligodendrocyte Cell-surface marker that characterizes mature oligodendrocyte Synaptophysin Neuron Neuronal protein located in synapses; indicates connections betweenneuronsTau Neuron Type of MAP; helps maintain structure of the axonPancreasCytokeratin 19 (CK19) Pancreatic epithelium CK19 identifies specific pancreatic epithelial cells that are progenitorsfor islet cells and ductal cellsGlucagon PancreaticisletExpressedby alpha-islet cell of pancreasInsulin Pancreatic islet Expressed by beta-islet cell of pancreas PancreasInsulin-promoting factor-1(PDX-1)Pancreatic islet Transcription factor expressed by beta-islet cell of pancreasNestin Pancreatic progenitor Structural filament protein indicative of progenitor cell lines includingpancreaticPancreatic polypeptide Pancreatic islet Expressed by gamma-islet cell of pancreasSomatostatin Pancreatic islet Expressed by delta-islet cell of pancreasPluripotent Stem CellsAlkaline phosphatase Embryonic stem (ES),embryonal carcinoma(EC) Elevated expression of this enzyme is associated with undifferentiated pluripotent stem cell (PSC)Alpha-fetoprotein (AFP) Endoderm Protein expressed during development of primitive endoderm; reflectsendodermal differentiation Pluripotent Stem CellsBone morphogenetic protein-4 Mesoderm Growth and differentiation factor expressed during early mesodermformation and differentiationBrachyury Mesoderm Transcription factor important in the earliest phases of mesodermformation and differentiation; used as the earliest indicator ofmesoderm formationCluster designation 30(CD30)ES, EC Surface receptor molecule found specifically on PSCCripto (TDGF-1) ES, cardiomyocyte Gene for growth factor expressed by ES cells, primitive ectoderm, anddeveloping cardiomyocyteGATA-4 gene Endoderm Expression increases as ES differentiates into endodermGCTM-2 ES, EC Antibody to a specific extracellular-matrix molecule that is synthesizedby undifferentiated PSCsGenesis ES, EC Transcription factor uniquely expressed by ES cells either in or duringthe undifferentiated state of PSCsGerm cell nuclear factor ES, EC Transcription factor expressed by PSCsHepatocyte nuclear factor-4(HNF-4)Endoderm Transcription factor expressed early in endoderm formationNestin Ectoderm, neural andpancreatic progenitor Intermediate filaments within cells; characteristic of primitive neuroectoderm formationNeuronal cell-adhesion molecule (N-CAM) Ectoderm Cell-surface molecule that promotes cell-cell interaction; indicatesprimitive neuroectoderm formationOct-4 ES, EC Transcription factor unique to PSCs; essential for establishment andmaintenance of undifferentiated PSCsPax6 Ectoderm Transcription factor expressed as ES cell differentiates intoneuroepitheliumStage-specific embryonic antigen-3 (SSEA-3) ES, EC Glycoprotein specifically expressed in early embryonic developmentand by undifferentiated PSCsStage-specific embryonic antigen-4 (SSEA-4) ES, EC Glycoprotein specifically expressed in early embryonic developmentand by undifferentiated PSCsStem cell factor (SCF or c-Kit ligand) ES, EC, HSC, MSC Membrane protein that enhances proliferation of ES and EC cells,hematopoietic stem cell (HSCs), and mesenchymal stem cells (MSCs);binds the receptor c-KitTelomerase ES, EC An enzyme uniquely associated with immortal cell lines; useful foridentifying undifferentiated PSCsTRA-1-60 ES, EC Antibody to a specific extracellular matrix molecule is synthesized byundifferentiated PSCsTRA-1-81 ES, EC Antibody to a specific extracellular matrix molecule normallysynthesized by undifferentiated PSCsVimentin Ectoderm, neural andpancreatic progenitor Intermediate filaments within cells; characteristic of primitive neuroectoderm formationSkeletal Muscle/Cardiac/Smooth MuscleMyoD and Pax7 Myoblast, myocyte Transcription factors that direct differentiation of myoblasts intomature myocytesMyogenin and MR4 Skeletal myocyte Secondary transcription factors required for differentiation ofmyoblasts from muscle stem cellsMyosin heavy chain Cardiomyocyte A component of structural and contractile protein found incardiomyocyteMyosin light chain Skeletal myocyte A component of structural and contractile protein found in skeletalmyocyte。