Protocol to dissociate healthy and infected murine- and hamster-derived lung tissue for single-cell transcriptome analysis

Pennitz, Peter; Goekeri, Cengiz; Trimpert, Jakob; Wyler, Emanuel; Ebenig, Aileen; Weissfuss, Chantal; Mühlebach, Michael D.; Witzenrath, Martin; Nouailles, Geraldine

doi:10.1016/j.xpro.2022.101957

Cited by 2 publications

(1 citation statement)

References 14 publications

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“…White blood cells were isolated from EDTA-blood as previously described; steps included red blood cell lysis and cell filtration before counting. Lung cells (caudal lobe) were isolated as previously described 26,60 ; steps included enzymatic digestion, mechanical dissociation and filtration before counting in trypan blue. Buffers contained 2 µg ml −1 actinomycin D to prevent de novo transcription during the procedures.…”

Section: Cell Isolation From Blood and Lungsmentioning

confidence: 99%

Live-attenuated vaccine sCPD9 elicits superior mucosal and systemic immunity to SARS-CoV-2 variants in hamsters

et al. 2023

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Vaccines play a critical role in combating the COVID-19 pandemic. Future control of the pandemic requires improved vaccines with high efficacy against newly emerging SARS-CoV-2 variants and the ability to reduce virus transmission. Here we compare immune responses and preclinical efficacy of the mRNA vaccine BNT162b2, the adenovirus-vectored spike vaccine Ad2-spike and the live-attenuated virus vaccine candidate sCPD9 in Syrian hamsters, using both homogeneous and heterologous vaccination regimens. Comparative vaccine efficacy was assessed by employing readouts from virus titrations to single-cell RNA sequencing. Our results show that sCPD9 vaccination elicited the most robust immunity, including rapid viral clearance, reduced tissue damage, fast differentiation of pre-plasmablasts, strong systemic and mucosal humoral responses, and rapid recall of memory T cells from lung tissue after challenge with heterologous SARS-CoV-2. Overall, our results demonstrate that live-attenuated vaccines offer advantages over currently available COVID-19 vaccines.

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Section: Cell Isolation From Blood and Lungsmentioning

confidence: 99%

Live-attenuated vaccine sCPD9 elicits superior mucosal and systemic immunity to SARS-CoV-2 variants in hamsters

et al. 2023

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Comparison of Lung Tissue‐Derived Extracellular Vesicles Using Multiple Dissociation Methods for Profiling Protein Biomarkers

Han,

Ye,

et al. 2024

Biotechnology Journal

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Extracellular vesicles (EVs) operate as chemical messengers that facilitate intercellular communication. Emerging evidence has demonstrated that lung tissue‐derived EVs play pivotal roles in pulmonary physiological processes and have potential as biomarkers and therapeutics for lung diseases. Multiple methods have been proposed for the isolation of lung tissue‐derived EVs. However, the effects of different tissue pre‐treatments on lung EV isolation and subsequent disease biomarker discovery have not yet been comprehensively investigated. In this study, we compared the physical characteristics, recovery yields, and protein compositions of EVs isolated from lung tissues using three methods based on different tissue dissociation principles. Methodologically, the beneficial roles of blood perfusion and gentle meshing were emphasized based on their impact on EV yield and purity. These results demonstrate that different methods enrich distinct subpopulations of EVs that exhibit significant differences in their protein cargo and surface properties. These disparities directly affect the diagnostic detection of marker proteins related to lung diseases, including lung tumors, asthma, and pulmonary fibrosis. Collectively, these findings highlight the variations in EV characteristics resulting from the applied approaches and offer compelling suggestions for guiding researchers in selecting a suitable isolation method based on downstream functional studies and clinical applications.

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Protocol to dissociate healthy and infected murine- and hamster-derived lung tissue for single-cell transcriptome analysis

Cited by 2 publications

References 14 publications

Live-attenuated vaccine sCPD9 elicits superior mucosal and systemic immunity to SARS-CoV-2 variants in hamsters

Live-attenuated vaccine sCPD9 elicits superior mucosal and systemic immunity to SARS-CoV-2 variants in hamsters

Comparison of Lung Tissue‐Derived Extracellular Vesicles Using Multiple Dissociation Methods for Profiling Protein Biomarkers

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