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用于HDAC抑制剂研究的原代急性髓系白血病细胞长期培养物的建立与特性分析

Establishment and Characterization of Long-Term Cultures Derived from Primary Acute Myeloid Leukemia Cells for HDAC Inhibitor Research.

作者信息

Romanski Annette, Bug Gesine

机构信息

Institute for Transfusion Medicine and Immunohematology, Goethe University Frankfurt am Main, Red Cross Blood Donor Service Baden-Württemberg-Hessen, Frankfurt, Germany.

Department of Medicine II, Hematology and Oncology, University Hospital Frankfurt, Goethe-University, Theodor-Stern-Kai 7, 60590, Frankfurt am Main, Germany.

出版信息

Methods Mol Biol. 2017;1510:127-148. doi: 10.1007/978-1-4939-6527-4_10.

Abstract

Histone deacetylase (HDAC) inhibitors are promising drugs. These agents lead to growth inhibition, cell cycle arrest, premature senescence, and apoptosis of malignant cells. Aim of our studies was to determine the efficacy of HDAC inhibitors on the clinically most relevant population of human leukemic progenitor cells in vitro. We here present stroma-free long-term cultures (LTC) of primary acute myeloid leukemia (AML) cells as a useful system for drug sensitivity testing in functional assays. AML-LTC are established by isolating mononuclear cells from peripheral blood samples of AML patients followed by selection of CD34 progenitor cells. AML-LTC cells can be maintained in liquid culture supplemented with cytokines and utilized for in vitro analyses to assess proliferation, apoptosis, expression of surface proteins or intracellular proteins and signal transduction, respectively.

摘要

组蛋白去乙酰化酶(HDAC)抑制剂是很有前景的药物。这些药物可导致恶性细胞生长抑制、细胞周期停滞、早衰和凋亡。我们研究的目的是在体外确定HDAC抑制剂对临床上最相关的人类白血病祖细胞群体的疗效。我们在此展示了原代急性髓系白血病(AML)细胞的无基质长期培养(LTC),作为功能测定中药物敏感性测试的有用系统。AML-LTC是通过从AML患者外周血样本中分离单核细胞,然后选择CD34祖细胞建立的。AML-LTC细胞可以在补充细胞因子的液体培养中维持,并分别用于体外分析,以评估增殖、凋亡、表面蛋白或细胞内蛋白的表达以及信号转导。

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