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idiMACS Separator

MidiMACS分离器

Company: Miltenyi Biotec
Catalog#: 130-042-302
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Primary Cultures from Human GH-secreting or Clinically Non-functioning Pituitary Adenomas
Author:
Date:
2018-04-05
[Abstract]  Pituitary adenomas are among the more frequent intracranial tumors usually treated with both surgical and pharmacological–based on somatostatin and dopamine agonists–approaches. Although mostly benign tumors, the occurrence of invasive behaviors is often detected resulting in poorer prognosis. The use of primary cultures from human pituitary adenomas represented a significant advancement in the knowledge of the mechanisms of their development and in the definition of the determinants of their pharmacological sensitivity. Moreover, recent studies identified also in pituitary adenomas putative tumor stem cells representing, according to the current hypothesis, the real cellular targets to eradicate most malignancies. In this protocol, we describe the procedure to establish primary cultures ... [摘要]  垂体腺瘤是更常见的颅内肿瘤之一,通常用基于生长抑素和多巴胺激动剂手术的手术和药物治疗。 虽然多为良性肿瘤,但侵入性行为的发生常常被检测到,导致预后较差。 来自人类垂体腺瘤的原代培养物的使用代表了对其发育机制的知识以及其药理敏感性决定因素的定义方面的显着进步。 此外,最近的研究也在垂体腺瘤中发现了假定的肿瘤干细胞,根据目前的假设,它代表了根除大多数恶性肿瘤的真实细胞靶标。 在这个协议中,我们描述了从人垂体腺瘤建立原代培养的程序,以及如何选择,体外扩增和表型鉴定推定的垂体腺瘤干细胞。

【背景】垂体腺瘤是最常见的颅内肿瘤之一(高达15%),横断面研究发现每100,000名居民中约有90例发病,其中绝大多数为30岁以上的成年人。大约10%的未经选择的垂体在尸检时进行了检查(即考虑到之前未诊断为垂体疾病的受试者的垂体)( ,Molitch,2017)。尽管通常为良性肿瘤,但垂体腺瘤的处理可因与激素分泌过多相关的临床综合征或发展以治疗抗性,高增殖率,快速复发和绒毛外侵袭为特征的侵袭行为而复杂化(Carreno等人,2017)。成年垂体干细胞的持续存在(Florio,2011)导致垂体腺瘤(以及可能的其他良性瘤形成)的发展可以源自具有干细胞特性(主要是自我更新和分化)的肿瘤细胞的亚群能力),正如已经建立的恶性固体和血液肿瘤一样。

最近的实验证据表明,癌症干细胞(CSC)范例也适用于人和小鼠垂体腺瘤(Donangelo等人,2014; ...

Differentiation of Myeloid-derived Suppressor Cells from Murine Bone Marrow and Their Co-culture with Splenic Dendritic Cells
Author:
Date:
2017-09-20
[Abstract]  Myeloid-derived suppressor cells (MDSCs) possess the ability to suppress the immune response, and to amplify the regulatory properties of other immune cells, i.e., dendritic cells. Here we describe a protocol in which MDSCs were differentiated from murine bone marrow cells, and CD11c+ dendritic cells were purified from murine spleens. MDSCs and CD11c dendritic cells can be co-cultured and the immunoregulatory phenotype of the MDSCs-conditioned dendritic cells could be assessed by means of a specific functional in vivo experiment, i.e., a skin test as a measure of the delayed-type hypersensitivity reaction toward a poorly immunogenic antigen. [摘要]  骨髓来源的抑制细胞(MDSCs)具有抑制免疫应答的能力,并扩增其他免疫细胞即树突状细胞的调节特性。 在这里,我们描述了MDSC与鼠骨髓细胞分化的方案,并且从鼠脾中纯化CD11c +树突状细胞。 可以共培养MDSC和CD11c树突状细胞,并且可以通过特定的功能体内实验来评估MDSCs条件树突细胞的免疫调节表型,即皮肤试验作为延迟型超敏反应的量度 抗免疫原性较差的抗原。
【背景】骨髓来源的抑制细胞(MDSCs)是由早期分化阶段的巨噬细胞,粒细胞,树突状细胞和骨髓细胞的前体组成的骨髓细胞组(Youn等人,2008),其在肿瘤的淋巴组织中大量积累感染性小鼠以及感染性疾病,败血症和创伤的小鼠。这些细胞的主要特征是它们以Ag特异性和/或非特异性方式抑制T细胞应答的能力。这些细胞现在被认为是负责肿瘤相关免疫缺陷的主要细胞类型之一;涉及MDSC介导的免疫抑制的主要因素包括Arg1的高表达(Marvel和Gabrilovich,2015)。精氨酸酶1(Arg1)和吲哚胺2,3-双加氧酶1(IDO1)分别是催化L-精氨酸(L-Arg)和L-色氨酸(L-Trp)降解的免疫调节酶,导致局部氨基酸剥夺。此外,与Arg1不同,IDO1在树突细胞(DC)中也具有非酶信号传导活性(Mondanelli等,2017)。除了其固有的免疫抑制活性外,MDSC还可能扩增其他免疫细胞的调节特性,特别是在肿瘤微环境中。虽然建立了MDSC-巨噬细胞相互作用的一些机制(Ugel等,2015),MDSCs和DCs之间的串扰仍然不清楚(Ostrand-Rosenberg等,2012);为弥补这一差距,我们已经制定了该方案,并且我们证明了Arg1 ...

Isolation of THY1+ Undifferentiated Spermatogonia from Mouse Postnatal Testes Using Magnetic-activated Cell Sorting (MACS)
Author:
Date:
2016-12-20
[Abstract]  In mammals, homeostasis of many tissues rely on a subpopulation of cells, referred to as stem cells, to sustain an appropriate number of undifferentiated and differentiated cells. Spermatogonial stem cells (SSCs) provide the fundamental cellular source for spermatogenesis and are responsible for the lifelong maintenance of the germline pool in testes throughout the reproductive lifespan of males. To gain insight into germline stem cell biology and develop strategies for infertility treatment, several germ cell isolation methods have been reported in order to acquire good quality and quantity of undifferentiated spermatogonia. Among them, magnetic-activated cell sorting (MACS) is an efficient cell isolation method that requires less time and less initial cell numbers to obtain an enriched ... [摘要]  在哺乳动物中,许多组织的体内平衡依赖于称为干细胞的细胞亚群,以维持适量的未分化细胞和分化细胞。精原细胞干细胞(SSC)为精子发生提供了基本的细胞来源,并且负责终生维持雄性生殖期间睾丸中的种系池。为了获得对种系干细胞生物学的了解并制定不孕症治疗策略,已经报道了几种生殖细胞分离方法,以获得良好质量和数量的未分化精原细胞。其中,磁激活细胞分选(MACS)是一种有效的细胞分离方法,其需要较少的时间和较少的初始细胞数以使用抗原 - 抗体反应来获得富集的细胞群体。胸腺细胞抗原1(THY1,CD90.2)被认为是小鼠新生儿和成年睾丸中未分化精原细胞的表面标志物。在这里,我们描述了一个用于分离来自小鼠睾丸的一周龄THY1 + 细胞和四周龄THY1 + 细胞的方案。分离过程包括三个步骤:睾丸收集和单细胞悬浮,使用生物素缀合的抗THY1抗体的细胞标记和磁性细胞分离。注意,这种隔离方案应在五小时内完成,以最大限度地提高活细胞的质量和数量。

背景 在几个成人组织中观察到活性和静止干细胞的共存。静止,自我更新和分化之间的充分平衡对于维持适当数量的未分化干细胞是必要的,并且避免过早的干细胞耗尽许多组织的体内平衡(Tseng等人,2015; Wabik和Jones,2015; ...

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