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Quantitative control of mitochondria transfer between live single cells using a microfluidic device

机译:使用微流控设备定量控制活单细胞之间线粒体的转移

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Quantitative control of mitochondria transfer between live cells is a promising approach for genetic manipulation of mitochondrial DNA (mtDNA) because single mitochondrion transfer to a mtDNA-less (ρ0) cell potentially leads to homoplasmy of mtDNA. In this paper, we describe a method for quantitative control of mitochondria transfer between live single cells. For this purpose, we fabricated novel microfluidic devices having cell paring structures with a 4.1, 5.6 or 10.0?μm-length microtunnel. When cells were fused through a microtunnel using the Sendai virus envelope-based method, a strictured cytoplasmic connection was achieved with a length corresponding to that of the microtunnel. Elongation of the cytoplasmic connection led to a decrease in mitochondria transfer to the fusion partner. Moreover, some cell pairs that fused through a 10.0?μm-length microtunnel showed single mitochondrion transfer. Fused cells were spontaneously disconnected from each other when they were recovered in a normal culture medium. These results suggest that our cell fusion method can perform quantitative control of mitochondria transfer that includes a single mitochondrion transfer.
机译:定量控制活细胞之间线粒体的转移是对线粒体DNA(mtDNA)进行遗传操作的一种有前途的方法,因为单个线粒体转移至无mtDNA(ρ0)的细胞可能导致mtDNA的同质。在本文中,我们描述了一种定量控制活单细胞之间线粒体转移的方法。为此目的,我们制造了新颖的微流控设备,该设备具有长度为4.1、5.6或10.0?m的微隧道的细胞配对结构。当使用基于仙台病毒包膜的方法通过微隧道融合细胞时,可以达到严格的胞质连接,其长度相当于微隧道的长度。细胞质连接的延长导致线粒体向融合伴侣的转移减少。此外,一些通过10.0?m长的微隧道融合的细胞对显示出单线粒体转移。当融合细胞在正常培养基中恢复时,它们会自发地彼此断开。这些结果表明,我们的细胞融合方法可以定量控制线粒体的转移,其中包括单个线粒体的转移。

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