Isolation of human melanoma stem cells using ALDH as a marker.
Isolation of human melanoma stem cells using ALDH as a marker.
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DOI：
10.1002/9780470151808.sc0308s26
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发表时间：
2013-09-20
影响因子：
--
通讯作者：
Fujita, Mayumi
中科院分区：
文献类型：
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作者：
Luo, Yuchun;Nguyen, Nicholas;Fujita, Mayumi
本单元描述了建立人类黑色素瘤患者来源的肿瘤异种移植模型（PDTX 模型）并使用乙醛脱氢酶 (ALDH) 作为标记物从人类黑色素瘤标本中分离人类黑色素瘤干细胞的方案。分析来自患者肿瘤样本的一小部分癌症干细胞的一个主要限制是无法获得大量的新鲜肿瘤组织。为了克服这一挑战，我们建立了人类黑色素瘤的 PDTX 模型。在该模型中，从黑色素瘤患者获得的人类肿瘤组织被解剖成小块，然后植入免疫功能低下的小鼠体内。 PDTX肿瘤保留了原始肿瘤的基本基因型和表型特征，适合进一步的生物学分析。使用 PDTX 模型，我们分析了 ALDH 标记的人类黑色素瘤干细胞。本单元将描述如何使用人类肿瘤样本建立PDTX模型。我们还将描述如何使用该模型分离和分析 ALDH 标记的人类黑色素瘤干细胞。
This unit describes a protocol for establishing a patient-derived tumor xenograft model (PDTX model) of human melanoma and isolating human melanoma stem cells from human melanoma specimens using aldehyde dehydrogenase (ALDH) as a marker. One major limitation of analyzing a small fraction of cancer stem cells from patient tumor samples is that substantial quantities of fresh tumor tissues are not available. To overcome this challenge, we have established a PDTX model of human melanoma. In this model, human tumor tissues obtained from melanoma patients are dissected into small pieces and subsequently implanted into immunocompromised mice. The PDTX tumors retain fundamental genotypic and phenotypic features of the original tumors and are suitable for further biological analyses. Using the PDTX model, we have analyzed ALDH-labeled human melanoma stem cells. This unit will describe how to establish a PDTX model using human tumor samples. We will also describe how to isolate and analyze ALDH-labeled human melanoma stem cells using this model.