Dielectrophoretic cell separation and gene expression profiling on microelectronic chip arrays
Dielectrophoretic cell separation and gene expression profiling on microelectronic chip arrays
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DOI：
10.1021/ac011273v
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发表时间：
2002-07-15
影响因子：
7.4
通讯作者：
Xu, X
中科院分区：
文献类型：
--
作者：
Huang, Y;Joo, S;Xu, X
通过 5 x 5 微电子芯片阵列上的介电泳交叉频率测量，确定了五种不同培养细胞系和人外周血单核细胞 (PBMC) 的细胞膜介电特性。基于各个细胞类型之间明显的介电特性差异，通过在该 5 x 5 阵列上进行介电电泳实现了有效的细胞分离，其中包括从 PBMC 中分离单核细胞 (U937) 或人 T 细胞白血病病毒 1 型 (HTLV-1) Tax 转化细胞 (Ind-2)，以及从神经胶质瘤细胞 (HTB) 中分离神经母细胞瘤细胞 (SH-SY5Y)。介电泳分离的细胞纯度可大于95%。通过电场促进杂交在 10 x 10 微电子芯片阵列上测定分离前后与 PBMC 混合的 U937 细胞的 IL-1、TNF-α 和 TGF-β 基因的表达谱。通过使用纯U937细胞的表达水平作为对照，表明分离后细胞的基因表达谱与分离前细胞混合物的基因表达谱显着不同。 U937 细胞在脂多糖诱导后基因表达水平的增加只能在分离后的细胞中准确测定，而分离前的样品掩盖了这些变化。此外，通过培养分离的HTB和SH-SY5Y细胞并测量应激相关基因c-fos的表达，结果表明介电泳力对细胞存活和应激几乎没有影响。所提出的使用微电子芯片阵列进行细胞分离和基因表达谱分析的方法为异质样品中特定细胞亚群的准确遗传分析提供了巨大的潜力。
Cell membrane dielectric properties of five different cultivated cell lines and human peripheral blood mononuclear cells (PBMC) were determined from dielectro-phoretic crossover frequency measurements on a 5 x 5 microelectronic chip array. Based on distinct dielectric property differences between individual cell types, efficient cell separations were achieved by dielectrophoresis on this 5 x 5 array, which included separation of monocytic cells (U937) or human T cell leukemia virus type 1 (HTLV-1) tax-transformed cells (Ind-2) from PBMC, as well as separation of nenroblastoma cells (SH-SY5Y) from glioma cells (HTB). The purity of dielectrophoretically separated cells can be greater than 95%. Expression profiles of IL-1, TNF-alpha, and TGF-beta genes for U937 cells mixed with PBMC before and after the separation were determined by a means of electric field-facilitated hybridization on a 10 x 10 microelectronic chip array. By using the expression levels of pure U937 cells as a control, it was shown that the gene expression profiles of the postseparation cells were significantly different from those of the preseparation cell mixtures. The increase in gene expression levels for U937 cells upon lipopolysacchride induction could be accurately determined only in the postseparation cells, while the preseparation samples masked these changes. Furthermore, by cultivating the separated HTB and SH-SY5Y cells and measuring expression of the stress-related gene c-fos, dielectrophoretic forces were shown to have little effect on cell survival and stress. The presented approach of using microelectronic chip arrays for both cell separation and gene expression profiling provides a great potential for accurate genetic analysis of specific cell subpopulations in heterogeneous samples.