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| Status |
Public on Apr 30, 2024 |
| Title |
Impact of different tissue dissociation protocols on endothelial cell recovery from developing lungs. |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Developing mouse lungs (day 14 of post-natal life) were dissociated by two different enzymatic methods, one employing commercially available Dispase, and another employing a commercial preparation of purified collagenase. The endothelial cell populations recovered from both approaches were characterized first by flow cytometry, to assess endothelial cell purity and viability. Subsequently, single-cell RNA-Seq was employed to characterize the constituent subpopulations of endothelial cells within each preparation. The single-cell RNA-Seq revealed that endothelial cell populations generated with Dispase exhibited reduced complexity, being enriched largely with one subpopulation of microvascular endothelial cells (gCap2) characterized by H2-Ab1 and Trf expression, whilst endothelial cell populations generated using collagenase yielded a complex endothelial cell population that included endothelial cell sub-types derived from the macro- and microvasculature, including the arterial and venous circulation, as well as the lymphatics.
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| Overall design |
Endothelial cells were isolated from mouse lungs on the 14th day of post-natal life. Lungs were cleared by vascular perfused, and enzymatically dissociated using either Dispase of collagenase. Endothelial cells were isolated by from the resultant single-cell suspensions using positive CD31 and negative CD45 selection. Life/dead cell discrimination was undertaken using DAPI exclusion by live cells. Resultant cell preparations were subjected to single-cell RNA-Seq analysis to allow clustering of constituent endothelial cell subtypes by principal component analysis.
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| Contributor(s) |
Palumbo F, Morty RE, Günther S |
| Citation(s) |
38668123 |
| Submission date |
Jul 07, 2023 |
| Last update date |
May 01, 2024 |
| Contact name |
Stefan Günther |
| E-mail(s) |
stefan.guenther@mpi-bn.mpg.de
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| Organization name |
MPI for heart and lung research
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| Street address |
ludwigtr. 43
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| City |
bad nauheim |
| ZIP/Postal code |
61231 |
| Country |
Germany |
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| Platforms (1) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA992552 |
