Microfluidic culture of single human embryonic stem cell colonies.
Microfluidic culture of single human embryonic stem cell colonies.
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DOI：
10.1039/b820380f
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发表时间：
2009-06-21
期刊：
影响因子：
6.1
通讯作者：
Smith GD
中科院分区：
文献类型：
--
作者：
Villa-Diaz LG;Torisawa YS;Uchida T;Ding J;Nogueira-de-Souza NC;O'Shea KS;Takayama S;Smith GD
我们已经开发了一种小型化的微流体培养系统，允许在动态（流动应用）或静态（无流动）条件下对单个人胚胎干细胞（hESC）集落进行实验。该系统由三个入口通道组成，这些入口通道汇聚成细胞培养通道，并通过使用模式化层流流体流向单个hESC集落的不同部分提供在空间和时间上递送特定治疗的能力。我们表明，微流控培养96小时或没有流动的结果在类似的维护hESC自我更新，分化成三个生殖细胞谱系的能力，并保持正常的核型，在标准培养皿。荧光核酸染料的局部递送是用层流实现的，仅在暴露细胞的细胞核中产生染色。同样，可以用酶处理去除集落的期望区域中的细胞，并收集用于分析。用细胞外基质重新包被通道的酶处理区域导致hESC集落重新生长到该区域中。我们的研究证明了在微流控装置中培养hESC，该装置可以将特定治疗递送到单个菌落的所需区域。这种小型化的培养系统允许原位处理和分析，具有从部分集落获得细胞样品而无需显微操作的能力，并且在允许hESC集落进一步生长的同时进行灵敏的分子分析。
We have developed a miniaturized microfluidic culture system that allows experimentation on individual human embryonic stem cell (hESC) colonies in dynamic (flow applied) or static (without flow) conditions. The system consists of three inlet channels that converge into a cell-culture channel and provides the capability to spatially and temporally deliver specific treatments by using patterned laminar fluid flow to different parts of a single hESC colony. We show that microfluidic culture for 96 h with or without flow results in similar maintenance of hESC self-renewal, the capability to differentiate into three germ cell lineages, and to maintain a normal karyotype, as in standard culture dishes. Localized delivery of a fluorescent nucleic acid dye was achieved with laminar flow, producing staining only in nuclei of exposed cells. Likewise, cells in desired regions of colonies could be removed with enzymatic treatment and collected for analysis. Re-coating the enzyme treated area of the channel with extracellular matrix led to re-growth of hESC colonies into this region. Our study demonstrates the culture of hESCs in a microfluidic device that can deliver specific treatments to desired regions of a single colony. This miniaturized culture system allows in situ treatment and analysis with the ability to obtain cell samples from part of a colony without micromanipulation and to perform sensitive molecular analysis while permitting further growth of the hESC colony.
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克隆衍生的人类胚胎干细胞系在长期培养过程中保持多能性和增殖潜力
影响因子：
2.7
作者：
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通讯作者：
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影响因子：
3.8
作者：
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影响因子：
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DOI：
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发表时间：
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影响因子：
11.1
作者：
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通讯作者：
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影响因子：
7.4
作者：
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通讯作者：
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