Identification of rare progenitor cells from human periosteal tissue using droplet microfluidics
Identification of rare progenitor cells from human periosteal tissue using droplet microfluidics
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DOI：
10.1039/b910472k
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发表时间：
2009-01-01
期刊：
影响因子：
4.2
通讯作者：
Edel, Joshua B.
中科院分区：
文献类型：
--
作者：
Srisa-Art, Monpichar;Bonzani, Ian C.;Edel, Joshua B.
从异质群体中分离和表征单细胞是细胞生物学、免疫学、干细胞研究和癌症研究中的重要过程。在新型细胞疗法的开发中，非常需要针对特定细胞类型，以便进行进一步的离体分析和扩增。我们在此介绍使用基于液滴的微流体作为平台技术来识别和量化不同的细胞表型。利用抗体和荧光染料对特定细胞群进行分子标记，可以使用高灵敏度共焦荧光检测来检测皮升大小的水滴中封装的单细胞。具体来说，在从人类骨膜组织中分离的异质细胞群中对稀有祖细胞进行了免疫检测。使用该模型人类细胞群，测试了液滴系统的准确性和再现性，并使用常规流式细胞术验证了结果。结果发现，使用两种技术测量的表型亚群的定量具有直接可比性。因此，这项研究证明了基于液滴的微流体用于细胞分析的生物能力，并为开发新型细胞分选技术提供了必要的第一步。
The isolation and characterisation of single cells from a heterogeneous population are important processes in cell biology, immunology, stem cell research, and cancer research. In the development of novel cell-based therapies, there is a considerable need to target specific cell types to allow for further analysis and amplification ex vivo. We introduce, herein, the use of droplet-based microfluidics as a platform technology for the identification and quantification of distinct cell phenotypes. Using molecular labelling of specific cell populations by antibodies and fluorescent dyes, detection of single cells encapsulated within picolitre-sized aqueous droplets can be performed using high-sensitivity confocal fluorescence detection. Specifically, rare progenitor cells were immunodetected within a heterogeneous population of cells isolated from human periosteal tissue. Using this model human cell population, the accuracy and reproducibility of the droplet system were tested and the results were verified using conventional flow cytometry. It was found that the quantitation of phenotypic subpopulations measured using both techniques is directly comparable. Accordingly, this study demonstrates the biological capacity of droplet-based microfluidics for cellular analysis and provides a necessary first step towards the development of a novel cell sorting technology.