Experiment Overview
| Repository ID: | FR-FCM-Z6C8 | Experiment name: | Human lung tissue digestion | MIFlowCyt score: | 50.75% |
| Primary researcher: | Mark Snyder | PI/manager: | Mark Snyder | Uploaded by: | Mark Snyder |
| Experiment dates: | 2021-05-12 - 2022-05-15 | Dataset uploaded: | May 2023 | Last updated: | Jun 2023 |
| Keywords: | [Immunophenotyping] [spectral flow cytometry] [Lung Tissue Processing] [Single cell RNA sequencing] | Manuscripts: | |||
| Organizations: |
University of Pittsburgh School of Medicine, Medicine, Pittsburgh, Pennsylvania (United States)
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| Purpose: | To determine the impact of different enzymatic digestion protocols on the cellular yield and phenotype from peripheral human lung tissue. | ||||
| Conclusion: | Protocols high in dispase cleaved immune markers CD4, CD8, CD69, and CD103 and contributed to an increased monocyte to macrophage yield. Similarly, dispase led to a differential epithelial cell yield, with increased TSPN8+ and ITGA6+ epithelial cells and reduced CD66e+ cells. When compared to collagenase D, collagenase P protocols yielded increased AT1 and AT2 cells and decreased endothelial cells. These results provide a framework for selecting an enzymatic digestion protocol best suited to the scientific question and allow for comparison of studies using different protocols. | ||||
| Comments: | Protocol 1: Mechanical digestion alone (gentleMACS) Protocol 2: Liberase + Dnase 1 (Disp lo) Protocol 3: Disp + Col D + Dnase 1 (Disp hi) Protocol 4: Col P + Dnase 1 (Col_P) Protocol 5: Col D + Dnase 1 + trypsin in (Col_D) Protocol 6: Col_D + density centrifugation | ||||
| Funding: | Not disclosed | ||||
| Quality control: | None | ||||
Experiment variables
| Conditions | |
|---|---|
| · Mechanical Digestion alone | 2020-026-C_Protocol_1.fcs · 2021-38-C_Protocol_1.fcs · 2021-39-C_Protocol_1.fcs · 2021-40-C_Protocol_1.fcs · 2021-41-C_Protocol_1.fcs |
| · Liperase TM | 2020-026-C_Protocol_2.fcs · 2021-38-C_Protocol_2.fcs · 2021-39-C_Protocol_2.fcs · 2021-40-C_Protocol_2.fcs · 2021-41-C_Protocol_2.fcs |
| · Dispase | 2020-026-C_Protocol_3.fcs · 2021-38-C_Protocol_3.fcs · 2021-39-C_Protocol_3.fcs · 2021-40-C_Protocol_3.fcs · 2021-41-C_Protocol_3.fcs |
| · Col_P | 2020-026-C_Protocol_4.fcs · 2021-38-C_Protocol_4.fcs · 2021-39-C_Protocol_4.fcs · 2021-40-C_Protocol_4.fcs · 2021-41-C_Protocol_4.fcs |
| · Col_D | 2020-026-C_Protocol_5.fcs · 2021-38-C_Protocol_5.fcs · 2021-39-C_Protocol_5.fcs · 2021-40-C_Protocol_5.fcs · 2021-41-C_Protocol_5.fcs |
| · Col_D+ | 2020-026-C_Protocol_6.fcs · 2021-38-C_Protocol_6.fcs · 2021-39-C_Protocol_6.fcs · 2021-40-C_Protocol_6.fcs · 2021-41-C_Protocol_6.fcs |
