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Label-free concentration of viable neurons, hESCs and cancer cells by means of acoustophoresis.

机译:通过声泳,无标记浓度的活神经元,hEsC和癌细胞。

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摘要

Concentration of viable cell populations in suspension is of interest for several clinical and pre-clinical applications. Here, we report that microfluidic acoustophoresis is an effective method to efficiently concentrate live and viable cells with high target purity without any need for protein fluorescent labeling using antibodies or over-expression. We explored the effect of the acoustic field acoustic energy density and systematically used different protocols to induce apoptosis or cell death and then determined the efficiency of live and dead cell separation. We used the breast cancer cell line MCF-7, the mouse neuroblastoma N2a as well as human embryonic stem cells (hESCs) to demonstrate that this method is gentle and can be applied to different cell populations. First, we induced cell death by means of high osmotic shock using a high concentration of PBS (10×), the protein kinase inhibitor staurosporine, high concentrations of dimethyl sulfoxide (DMSO, 10%), and finally, cell starvation. In all the methods employed, we successfully induced cell death and were able to purify and concentrate the remaining live cells using acoustophoresis. Importantly, the concentration of viable cells was not dependent on a specific cell type. Further, we demonstrate that different death inducing stimuli have different effects on the intrinsic cell properties and therefore affect the efficiency of the acoustophoretic separation.
机译:对于几种临床和临床前应用而言,悬浮中活细胞群体的浓度是令人感兴趣的。在这里,我们报道微流声电泳是一种有效的方法,可以有效地浓缩具有高目标纯度的活细胞和活细胞,而无需使用抗体或过度表达进行蛋白质荧光标记。我们探索了声场声能密度的影响,系统地使用了不同的方案来诱导细胞凋亡或细胞死亡,然后确定了活细胞和死细胞分离的效率。我们使用了乳腺癌细胞系MCF-7,小鼠神经母细胞瘤N2a以及人类胚胎干细胞(hESCs)来证明这种方法是温和的,可以应用于不同的细胞群。首先,我们通过使用高浓度的PBS(10x),蛋白激酶抑制剂星形孢菌素,高浓度的二甲基亚砜(DMSO,10%)进行高渗透压休克诱导细胞死亡,最后是细胞饥饿。在所有使用的方法中,我们成功地诱导了细胞死亡,并能够通过声泳纯化和浓缩剩余的活细胞。重要的是,活细胞的浓度不取决于特定的细胞类型。此外,我们证明了不同的死亡诱导刺激对固有细胞特性具有不同的影响,因此影响了声电泳分离的效率。

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