The Construction and Identification of Induced Pluripotent Stem Cells Derived from Acute Myelogenous Leukemia Cells
The Construction and Identification of Induced Pluripotent Stem Cells Derived from Acute Myelogenous Leukemia Cells
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DOI：
10.1159/000471246
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发表时间：
2017-03
影响因子：
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通讯作者：
Liang-fang Zhu;Qiuru Chen;Shaozhen Chen;Ling-yan Wang;Xiao-feng Luo;Jin-Hua Ren;Xiaohong Yuan;Xueqiong Wu;Yanling Zeng;Min Xiao;Yongquan Chen;Y. Chen;Minhui Lin;Zhengjun Wu;Zhi-zhe Chen;Jian-da Hu;Ting Yang
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文献类型：
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作者：
Liang-fang Zhu;Qiuru Chen;Shaozhen Chen;Ling-yan Wang;Xiao-feng Luo;Jin-Hua Ren;Xiaohong Yuan;Xueqiong Wu;Yanling Zeng;Min Xiao;Yongquan Chen;Y. Chen;Minhui Lin;Zhengjun Wu;Zhi-zhe Chen;Jian-da Hu;Ting Yang
目的：本研究旨在从急性髓性白血病（AML）细胞中建立诱导多能干细胞（iPSC）系，并鉴定其生物学特性。方法：用携带OCT 4、SOX 2、KLF 4和C-MYC的慢病毒感染来自M6患者的AML浸润皮肤的细胞以诱导iPSCs。通过碱性磷酸酶（ALP）染色证实iPSC的特征。CCK-8法检测iPSCs的增殖能力。免疫组化检测多能性标志物的表达; qRT-PCR检测干细胞相关基因的表达;核型分析检测诱导过程中的畸变;拟胚体形成和畸胎瘤形成试验检测iPSCs的分化潜能。ALP染色证实这些细胞呈阳性染色，具有iPSCs的特征。结果：CCK-8法显示诱导的iPSCs具有增殖能力。免疫染色显示iPSC克隆显示NANOG、SSEA-3、SSEA-4、TRA-1-60和TRA-1-81的阳性表达。qRT-PCR结果显示Nanog、Lin 28、Cripto、FOX 3、DNMT 3b、DPPA 2和DPPA 4的mRNA表达在iPSCs中显著增加。核型分析未发现iPSC中的染色体畸变。胚状体形成和畸胎瘤形成测定的结果表明，iPSC具有分化成所有三个胚层的潜力。结论：本研究成功建立了AML细胞诱导的iPSC细胞系。
Objective: The present study aimed to establish an induced pluripotent stem cell (iPSC) line from acute myelogenous leukemia (AML) cells in vitro and identify their biological characteristics. Methods: Cells from the AML-infiltrated skin from an M6 patient were infected with a lentivirus carrying OCT4, SOX2, KLF4 and C-MYC to induce iPSCs. The characteristics of the iPSCs were confirmed by alkaline phosphatase (ALP) staining. The proliferation ability of iPSCs was detected with a CCK-8 assay. The expression of pluripotency markers was measured by immunostaining, and the expression of stem cell-related genes was detected by qRT-PCR; distortion during the induction process was detected by karyotype analysis; the differentiation potential of iPSCs was determined by embryoid body-formation and teratoma-formation assays. ALP staining confirmed that these cells exhibited positive staining and had the characteristics of iPSCs. Results: The CCK-8 assay showed that the iPSCs had the ability to proliferate. Immunostaining demonstrated that iPSC clones showed positive expression of NANOG, SSEA-3, SSEA-4, TRA-1-60 and TRA-1-81. qRT-PCR results revealed that the mRNA expression of Nanog, Lin28, Cripto, FOX3, DNMT3b, DPPA2, and DPPA4 significantly increased in iPSCs. Karyotype analysis found no chromosome aberration in the iPSCs. The results of the embryoid body-formation and teratoma-formation assays indicated that the iPSCs had the potential to differentiate into all three germ layers. Conclusion: Our study provided evidence that an iPSC line derived from AML cells was successfully established.