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| Status |
Public on Jan 25, 2022 |
| Title |
Single-cell transcriptomics of dynamic cell behaviors |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Despite advances in spatial transcriptomics, linking dynamic behavior of cells in their native environment to molecular profiling remains a major challenge. We present a method, termed behavioral transcriptomics, that allows us to couple physiological behaviors of single cells in an intact tissue to deep molecular profiling of individual cells. This method enables us to establish a novel molecular signature for a striking migratory cellular behavior following tissue injury.
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| Overall design |
In order to visualize the airway epithelium at high resolution over days, we explant a mouse trachea and secure it in a custom imaging platform, which minimizes sample movement during imaging and maintains a constant supply of nutrients from below the explant without disrupting the air-liquid interface. To determine the molecular signatures of cells with directional movement compared to regenerating non-moving cells, we marked epithelial regions by photoconversion at 24 hours after injury, imaged every 6 hours, screened for “movers” with >50 μm displacement (>3 μm/hr) at 18 hours after photoconversion, and then isolated photoconverted epithelial cells for plate-based single cell sequencing (scSeq). Dimensionality reduction revealed that cells from “moving” region (M) and a “non-moving” (NM) region cluster separately.
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| Contributor(s) |
Kwok SJ, Montoro DT, Haber AL, Yun S, Vinarsky V |
| Citation(s) |
37083555 |
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| Submission date |
Jan 19, 2022 |
| Last update date |
Aug 04, 2023 |
| Contact name |
Adam Haber |
| E-mail(s) |
ahaber@hsph.harvard.edu
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| Organization name |
Harvard T. H. Chan School of Public Health
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| Street address |
665 Huntington Ave, HSPH Building 1, Room 305
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| City |
Boston |
| State/province |
Massachusetts |
| ZIP/Postal code |
02115 |
| Country |
USA |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (131)
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| Relations |
| BioProject |
PRJNA798600 |
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