Dissociation and inDrops microfluidic encapsulation of human gut tissues for single-cell atlasing studies
Summary: In droplet-based single-cell RNA-sequencing (scRNA-seq) experiments, cells, along with some of their surrounding buffer and ambient material, are encapsulated into droplets for mRNA capture and barcoding. This protocol details the steps for human gut tissue dissociation using cold active pr...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722004506 |
| _version_ | 1817970531480109056 |
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| author | Alan J. Simmons Ken S. Lau |
| author_facet | Alan J. Simmons Ken S. Lau |
| author_sort | Alan J. Simmons |
| collection | DOAJ |
| description | Summary: In droplet-based single-cell RNA-sequencing (scRNA-seq) experiments, cells, along with some of their surrounding buffer and ambient material, are encapsulated into droplets for mRNA capture and barcoding. This protocol details the steps for human gut tissue dissociation using cold active protease, and subsequent isolation of single epithelial cells, with enrichment of viability through washes. Next, the steps for encapsulation on the inDrops scRNA-seq platform are described. This procedure has been demonstrated to be applicable to polyps, cancers, and inflamed tissues.For complete details on the use and execution of this protocol, please refer to Chen et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-13T20:35:33Z |
| format | Article |
| id | doaj.art-8c683301736c427195744722e5bb196a |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-13T20:35:33Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-8c683301736c427195744722e5bb196a2022-12-22T02:31:03ZengElsevierSTAR Protocols2666-16672022-09-0133101570Dissociation and inDrops microfluidic encapsulation of human gut tissues for single-cell atlasing studiesAlan J. Simmons0Ken S. Lau1Epithelial Biology Center and Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Corresponding authorEpithelial Biology Center and Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Corresponding authorSummary: In droplet-based single-cell RNA-sequencing (scRNA-seq) experiments, cells, along with some of their surrounding buffer and ambient material, are encapsulated into droplets for mRNA capture and barcoding. This protocol details the steps for human gut tissue dissociation using cold active protease, and subsequent isolation of single epithelial cells, with enrichment of viability through washes. Next, the steps for encapsulation on the inDrops scRNA-seq platform are described. This procedure has been demonstrated to be applicable to polyps, cancers, and inflamed tissues.For complete details on the use and execution of this protocol, please refer to Chen et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722004506Single CellRNAseq |
| spellingShingle | Alan J. Simmons Ken S. Lau Dissociation and inDrops microfluidic encapsulation of human gut tissues for single-cell atlasing studies STAR Protocols Single Cell RNAseq |
| title | Dissociation and inDrops microfluidic encapsulation of human gut tissues for single-cell atlasing studies |
| title_full | Dissociation and inDrops microfluidic encapsulation of human gut tissues for single-cell atlasing studies |
| title_fullStr | Dissociation and inDrops microfluidic encapsulation of human gut tissues for single-cell atlasing studies |
| title_full_unstemmed | Dissociation and inDrops microfluidic encapsulation of human gut tissues for single-cell atlasing studies |
| title_short | Dissociation and inDrops microfluidic encapsulation of human gut tissues for single-cell atlasing studies |
| title_sort | dissociation and indrops microfluidic encapsulation of human gut tissues for single cell atlasing studies |
| topic | Single Cell RNAseq |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722004506 |
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