ArticleJournal of clinical laboratory analysis2026
Standardization of the Monocyte Monolayer Assay: Lessons Learned From Laboratory Optimization and Literature Review.
Article in Journal of clinical laboratory analysis, 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
objectivesSeveral variables influence the outcome of cellular assays. We conducted a comprehensive literature review to identify the factors affecting the Monocyte Monolayer Assay (MMA) and subsequently standardized the assay through systematic in-house laboratory trials.
methodsThe study was conducted in two phases. Phase 1 involved a literature review using PubMed and Scopus databases to identify factors influencing the assay. Phase 2 included experimental trials to evaluate the effects of these factors. Monocytes were isolated from buffy coat bags, and assay parameters, such as centrifugation speed, monocyte adherence conditions, RBC sensitization, incubation time, and staining, were systematically standardized. A blinded evaluation was employed for slide reading to assess the influence of procedural factors on assay reliability.
resultsA literature review using the PubMed and Scopus databases identified key variables influencing MMA performance. Twelve sequential trials were conducted to identify the effects of these variables on assay outcome. Optimal centrifugation speed and time was 600 × g for 15 min, and the use of four buffy coat bags improved monocyte yield to 8.3 × 10
conclusionThis study emphasizes the importance of standardizing the MMA procedure before implementation. Optimizing the parameters to ensure enhanced monocyte yield, reduced cell lysis, and reliable interpretation for assessing the clinical significance of red cell alloantibodies is essential.
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