期刊
SMALL
卷 15, 期 5, 页码 -出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/smll.201804576
关键词
hydrogels; microdroplets; pluripotency; single cell analysis; stem cells
类别
资金
- Wellcome Trust [WT108438/C/15/Z]
- Engineering and Physical Sciences Research Council (EPSRC)
- EU via its Horizon2020 programme
- Medical Research Council (MRC)
- MRC
- Studienstiftung des deutschen Volkes
- German Academic Exchange Service (DAAD)
Developmental cell biology requires technologies in which the fate of single cells is followed over extended time periods, to monitor and understand the processes of self-renewal, differentiation, and reprogramming. A workflow is presented, in which single cells are encapsulated into droplets (empty set: 80 mu m, volume: approximate to 270 pL) and the droplet compartment is later converted to a hydrogel bead. After on-chip de-emulsification by electrocoalescence, these 3D scaffolds are subsequently arrayed on a chip for long-term perfusion culture to facilitate continuous cell imaging over 68 h. Here, the response of murine embryonic stem cells to different growth media, 2i and N2B27, is studied, showing that the exit from pluripotency can be monitored by fluorescence time-lapse microscopy, by immunostaining and by reverse-transcription and quantitative PCR (RT-qPCR). The defined 3D environment emulates the natural context of cell growth (e.g., in tissue) and enables the study of cell development in various matrices. The large scale of cell cultivation (in 2000 beads in parallel) may reveal infrequent events that remain undetected in lower throughput or ensemble studies. This platform will help to gain qualitative and quantitative mechanistic insight into the role of external factors on cell behavior.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据