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- Title
Droplet Microfluidics Enables Tracing of Target Cells at the Single-Cell Transcriptome Resolution.
- Authors
Liu, Yang; Wang, Shiyu; Lyu, Menghua; Xie, Run; Guo, Weijin; He, Ying; Shi, Xuyang; Wang, Yang; Qi, Jingyu; Zhu, Qianqian; Zhang, Hui; Luo, Tao; Chen, Huaying; Zhu, Yonggang; Dong, Xuan; Li, Zida; Gu, Ying; Liu, Longqi; Xu, Xun; Liu, Ya
- Abstract
The rapid promotion of single-cell omics in various fields has begun to help solve many problems encountered in research, including precision medicine, prenatal diagnosis, and embryo development. Meanwhile, single-cell techniques are also constantly updated with increasing demand. For some specific target cells, the workflow from droplet screening to single-cell sequencing is a preferred option and should reduce the impact of operation steps, such as demulsification and cell recovery. We developed an all-in-droplet method integrating cell encapsulation, target sorting, droplet picoinjection, and single-cell transcriptome profiling on chips to achieve labor-saving monitoring of TCR-T cells. As a proof of concept, in this research, TCR-T cells were encapsulated, sorted, and performed single-cell transcriptome sequencing (scRNA-seq) by injecting reagents into droplets. It avoided the tedious operation of droplet breakage and re-encapsulation between droplet sorting and scRNA-seq. Moreover, convenient device operation will accelerate the progress of chip marketization. The strategy achieved an excellent recovery performance of single-cell transcriptome with a median gene number over 4000 and a cross-contamination rate of 8.2 ± 2%. Furthermore, this strategy allows us to develop a device with high integrability to monitor infused TCR-T cells, which will promote the development of adoptive T cell immunotherapy and their clinical application.
- Subjects
TRANSCRIPTOMES; MICROFLUIDICS; T cells; INDIVIDUALIZED medicine; PRENATAL diagnosis
- Publication
Bioengineering (Basel), 2022, Vol 9, Issue 11, p674
- ISSN
2306-5354
- Publication type
Article
- DOI
10.3390/bioengineering9110674