ArticleScientific reports2026
Refinement of renal T lymphocytes enrichment strategies: advancing the in-depth study of renal immune responses.
Article in Scientific reports, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
The kidney T lymphocytes emerge as central agents in both physiological surveillance and pathological processes. In this study, we aimed to establish an optimized protocol for isolating and purifying T lymphocytes from mouse and human kidneys to provide support for renal diseases research. To develop an optimized protocol, we evaluated different tissue fragmentation strategies, enzymatic digestion conditions, digestion times and Percoll density gradients to maximize cell yield and viability while preserving surface markers. Subsequently, we applied this method to various kidney disease models and performed single-cell sequencing analysis by using public database. Our analyses revealed mechanical mincing combined with 0.2% collagenase IV and 0.02% DNase I for 45 min, followed by 33-80% Percoll gradient centrifugation, yielded the highest number of viable mononuclear cells with preserved surface markers. This protocol outperformed mechanical grinding and high concentration collagenase digestion, which caused greater mechanical and chemical damage. Flow cytometry confirmed optimal isolation of CD4
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