Multisite Assessment of Methods for Cell Preservation Upstream of Single-Cell RNA Sequencing
Rattachement africain : us. Niveau de preuve : code pays fourni par la source.
Le résumé fourni par la source
Introduction/Objective: Single-cell RNA sequencing (scRNA-seq) resolves cell types and molecular phenotypes within heterogeneous specimens but typically requires fresh, high-quality single-cell suspensions processed immediately to preserve transcriptional profiles. This constraint complicates samples with long preparation times and prevents collection at remote sites lacking single-cell instrumentation. Several commercial assays now enable preservation at the point of collection through fixation or cryopreservation, allowing processing to occur months later. The Association of Biomolecular Research Facilities' DNA Sequencing and Genomics and Bioinformatics Research Groups undertook a cross-platform, multisite study to assess the performance and reproducibility of three such platforms: 10x Genomics FLEX, Parse Biosciences Evercode WT v2, and Honeycomb Bio HIVE. Materials and Methods: Total leukocytes and peripheral blood mononuclear cells (PBMCs) were isolated from a single healthy individual, with EasySep reagent used for red blood cell depletion of the leukocyte fraction. Cells were characterized by a 21-color flow cytometry panel to provide a reference, and the remaining material was fixed or cryopreserved according to each platform's protocol. Preserved leukocyte samples were prepared in parallel by two technicians ("A" and "B" replicates) and distributed to multiple ABRF member core facilities for downstream processing, while fresh leukocytes processed with the 10x 3' v3.1 (3pGEX) chemistry served as a reference. Libraries were sequenced at a central site, and performance was evaluated across standard scRNA-seq quality control metrics, gene and transcript detection sensitivity, cell-type discovery and annotation, differential expression, and correlation analyses. Results: Data from all platforms integrated effectively and produced concordant results for cell-type annotation and relative abundance, with cell-type proportions broadly consistent with the flow cytometry reference. However, platform-specific expression signatures were evident for a subset of genes, and cross-site reproducibility varied between methods, with the FLEX workflow showing greater susceptibility to technical variation introduced during on-site sample processing. Preservation-based methods (FLEX and HIVE) showed better retention of fragile granulocyte populations than fresh samples processed with the 10x 3pGEX workflow. Discussion: Improvements to preservation methods are changing how single-cell research is conducted by decoupling sample collection from downstream processing. Our investigation into the performance and reproducibility of each platform provides a resource to help investigators and core facilities select the most appropriate single-cell preservation workflow given their sample type, cell populations of interest, sample collection logistics, and laboratory infrastructure constraints.
Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.
Le contrôle bibliographique ouvert
DOI retrouvé dans Crossref DOI retrouvé ; titre concordant.
- Titre Crossref
- Multisite Assessment of Methods for Cell Preservation Upstream of Single-Cell RNA Sequencing
- Date Crossref
- 08/06/2026
- Éditeur
- Association of Biomolecular Resource Facilities
- Type
- journal-article
Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.
Où se fait cette recherche
-
Dartmouth College pays non établi dans la noticeUniversité ou école supérieure
-
Dartmouth Institute for Health Policy and Clinical Practice pays non établi dans la noticeÉtablissement de santé
-
Dartmouth Psychiatric Research Center pays non établi dans la noticeStructure de recherche
-
Dartmouth–Hitchcock Medical Center pays non établi dans la noticeÉtablissement de santé
-
Barrow Neurological Institute Flow Cytometry and Genomics Core Facility pays non établi dans la noticeÉtablissement de santé
-
University of Delaware Department of Computer and Information Sciences pays non établi dans la noticeUniversité ou école supérieure
-
Dana-Farber Cancer Institute pays non établi dans la noticeStructure de recherche
-
University at Albany Center for Functional Genomics and Department of Biomedical Sciences pays non établi dans la noticeUniversité ou école supérieure
-
Washington University in St. Louis pays non établi dans la noticeUniversité ou école supérieure
-
The University of Texas at San Antonio Genomic Sequencing and Analysis Facility pays non établi dans la noticeUniversité ou école supérieure
-
Indiana University – Purdue University Indianapolis Bioinformatics Core pays non établi dans la noticeUniversité ou école supérieure
-
University of Wisconsin–Madison pays non établi dans la noticeUniversité ou école supérieure
Dartmouth College, Dartmouth Institute for Health Policy and Clinical Practice et Dartmouth Psychiatric Research Center, avec 9 autres affiliations.
Une affiliation ne permet pas de déduire la nationalité d’un auteur.