Protocols for cytogenetic studies of human embryonic stem cells
Protocols for cytogenetic studies of human embryonic stem cells
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DOI：
10.1016/j.ymeth.2008.03.005
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发表时间：
2008-06-01
期刊：
影响因子：
4.8
通讯作者：
Johnson, Julie A.
中科院分区：
文献类型：
--
作者：
Meisner, Lorraine Faxon;Johnson, Julie A.
所有培养的细胞都会随着时间的推移发生染色体变化，包括人类胚胎干细胞（hESC）的培养物，但只有那些具有适应性染色体变化的细胞才能存活。hESC培养物中最常见的染色体变化是12三体和17三体。具有这些三体的细胞在外观上与正常细胞无法区分，并且还表现出多能性的典型标志物，使得它们在没有细胞遗传学分析的情况下难以识别。这些细胞的早期检测是必不可少的，因为具有12和17三体的细胞可以在5-10代中取代正常细胞群。使用G显带的细胞遗传学分析被认为是检测染色体异常的金标准，并且当与间期FISH结合使用时，提供了早期检测细胞遗传学畸变的灵敏方法，例如染色体12和17的完全和部分三体。以下讨论描述了我们实验室用于研究培养的hESC的细胞遗传学方法，沿着将这些方法整合到常规细胞系质量控制计划中的建议。(c)2008年爱思唯尔公司All rights reserved.
All cultured cells develop chromosome changes over time, including cultures of human embryonic stem cells (hESC), but only those cells with adaptive chromosomes changes survive. The most frequent chromosome changes in hESC cultures are trisomy 12 and trisomy 17. Cells with these trisomies are indistinguishable from normal cells by appearance and also demonstrate typical markers of pluripotency, making them difficult to identify without cytogenetic analysis. Early detection of these cells is essential since cells with trisomy 12 and 17 can replace the normal cell population in 5-10 passages. Cytogenetic analysis using G-banding is considered to be the gold standard for detecting chromosome abnormalities and, when used in combination with interphase FISH, provides a sensitive method for early detection of cytogenetic aberrations, such as full and partial trisomies of chromosomes 12 and 17. The following discussion describes the cytogenetic methods used in our laboratory to study cultured hESCs, along with recommendations for integrating these methods into a plan for routine cell line quality control. (c) 2008 Elsevier Inc. All rights reserved.