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使用醛脱氢酶（ALDH）作为标志物分离人黑色素瘤干细胞。

Isolation of human melanoma stem cells using ALDH as a marker.

作者信息

Luo Yuchun, Nguyen Nicholas, Fujita Mayumi

机构信息

Department of Dermatology, University of Colorado Denver, Aurora, Colorado.

Charles C. Gates Center for Regenerative Medicine and Stem Cell Biology, University of Colorado Denver, Aurora, Colorado.

出版信息

Curr Protoc Stem Cell Biol. 2013 Sep 20;26:3.8.1-3.8.10. doi: 10.1002/9780470151808.sc0308s26.

DOI:10.1002/9780470151808.sc0308s26
PMID:24510792
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4164114/
Abstract

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.

摘要

本单元介绍了一种建立人黑色素瘤患者来源肿瘤异种移植模型（PDTX模型）的方案，并使用醛脱氢酶（ALDH）作为标志物从人黑色素瘤标本中分离人黑色素瘤干细胞。从患者肿瘤样本中分析一小部分癌症干细胞的一个主要限制是无法获得大量新鲜肿瘤组织。为了克服这一挑战，我们建立了人黑色素瘤的PDTX模型。在该模型中，将从黑色素瘤患者获得的人肿瘤组织切成小块，随后植入免疫缺陷小鼠体内。PDTX肿瘤保留了原始肿瘤的基本基因型和表型特征，适用于进一步的生物学分析。利用PDTX模型，我们分析了ALDH标记的人黑色素瘤干细胞。本单元将描述如何使用人肿瘤样本建立PDTX模型。我们还将描述如何使用该模型分离和分析ALDH标记的人黑色素瘤干细胞。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f28/4164114/93a4946020f2/nihms-627223-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f28/4164114/79809d2e41c0/nihms-627223-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f28/4164114/3068077c4f89/nihms-627223-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f28/4164114/93a4946020f2/nihms-627223-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f28/4164114/79809d2e41c0/nihms-627223-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f28/4164114/3068077c4f89/nihms-627223-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f28/4164114/93a4946020f2/nihms-627223-f0003.jpg

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Stem Cells. 2012 Oct;30(10):2100-13. doi: 10.1002/stem.1193.
2
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Stem Cells. 2012 Sep;30(9):1808-18. doi: 10.1002/stem.1160.
3
Side population cells from human melanoma tumors reveal diverse mechanisms for chemoresistance.人黑色素瘤肿瘤中的侧群细胞揭示了多种化学抗性的机制。
J Invest Dermatol. 2012 Oct;132(10):2440-2450. doi: 10.1038/jid.2012.161. Epub 2012 May 24.
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Patient-derived tumour xenografts as models for oncology drug development.患者来源肿瘤异种移植模型在肿瘤药物开发中的应用。
Nat Rev Clin Oncol. 2012 Apr 17;9(6):338-50. doi: 10.1038/nrclinonc.2012.61.
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Human CD271-positive melanoma stem cells associated with metastasis establish tumor heterogeneity and long-term growth.人 CD271 阳性黑色素瘤干细胞与转移相关，可建立肿瘤异质性和长期生长。
Cancer Res. 2011 Apr 15;71(8):3098-109. doi: 10.1158/0008-5472.CAN-10-3997. Epub 2011 Mar 10.
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Phenotypic heterogeneity among tumorigenic melanoma cells from patients that is reversible and not hierarchically organized.肿瘤发生性黑色素瘤细胞在患者之间存在表型异质性，这种异质性是可逆的，且不是层次组织的。
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