Evidence map›Paper›PMID 29359175›Full record

ArticleNeurology(R) neuroimmunology & neuroinflammation2018

Defining standard enzymatic dissociation methods for individual brains and spinal cords in EAE.

Rehana Z Hussain, William A Miller-Little, Richard Doelger, Gary R Cutter, Nicolas Loof, Petra D Cravens, Olaf Stüve

Abstract read
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Article in Neurology(R) neuroimmunology & neuroinflammation, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.

0numbers the graph read from it
0cells of the map it votes in
11citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

11 citing papers in PubMed.

  1. Article
  2. Article
  3. Article
  4. Review
  5. CD11cProceedings of the National Academy of Sciences of the United States of America · 2021
    Article
  6. Article
  7. Preparing a Single Cell Suspension from Zebrafish Retinal Tissue for Flow Cytometric Cell Sorting of Müller Glia.Cytometry. Part A : the journal of the International Society for Analytical Cytology · 2020
    Article
  8. Article
  9. Article
  10. TLR3 agonism re-establishes CNS immune competence duringAnnals of clinical and translational neurology · 2018
    Article
  11. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors.

Rehana Z HussainDepartment of Neurology and Neurotherapeutics (R.Z.H., W.A.M.-L., R.D., P.D.C., O.S.), University of Texas Southwestern Medical Center, Dallas; Department of Biostatistics (G.C.), University of Alabama at Birmingham; The Moody Foundation Flow Cytometry Facility (N.L.), Children's Research Institute, University of Texas Southwestern Medical Center, Dallas; Neurology Section (O.S.), VA North Texas Health Care System, Medical Service, Dallas, TX; and Department of Neurology (O.S.), Klinikum rechts der Isar, Technische Universität München, Germany.
William A Miller-LittleDepartment of Neurology and Neurotherapeutics (R.Z.H., W.A.M.-L., R.D., P.D.C., O.S.), University of Texas Southwestern Medical Center, Dallas; Department of Biostatistics (G.C.), University of Alabama at Birmingham; The Moody Foundation Flow Cytometry Facility (N.L.), Children's Research Institute, University of Texas Southwestern Medical Center, Dallas; Neurology Section (O.S.), VA North Texas Health Care System, Medical Service, Dallas, TX; and Department of Neurology (O.S.), Klinikum rechts der Isar, Technische Universität München, Germany.
Richard DoelgerDepartment of Neurology and Neurotherapeutics (R.Z.H., W.A.M.-L., R.D., P.D.C., O.S.), University of Texas Southwestern Medical Center, Dallas; Department of Biostatistics (G.C.), University of Alabama at Birmingham; The Moody Foundation Flow Cytometry Facility (N.L.), Children's Research Institute, University of Texas Southwestern Medical Center, Dallas; Neurology Section (O.S.), VA North Texas Health Care System, Medical Service, Dallas, TX; and Department of Neurology (O.S.), Klinikum rechts der Isar, Technische Universität München, Germany.
Gary R CutterDepartment of Neurology and Neurotherapeutics (R.Z.H., W.A.M.-L., R.D., P.D.C., O.S.), University of Texas Southwestern Medical Center, Dallas; Department of Biostatistics (G.C.), University of Alabama at Birmingham; The Moody Foundation Flow Cytometry Facility (N.L.), Children's Research Institute, University of Texas Southwestern Medical Center, Dallas; Neurology Section (O.S.), VA North Texas Health Care System, Medical Service, Dallas, TX; and Department of Neurology (O.S.), Klinikum rechts der Isar, Technische Universität München, Germany.
Nicolas LoofDepartment of Neurology and Neurotherapeutics (R.Z.H., W.A.M.-L., R.D., P.D.C., O.S.), University of Texas Southwestern Medical Center, Dallas; Department of Biostatistics (G.C.), University of Alabama at Birmingham; The Moody Foundation Flow Cytometry Facility (N.L.), Children's Research Institute, University of Texas Southwestern Medical Center, Dallas; Neurology Section (O.S.), VA North Texas Health Care System, Medical Service, Dallas, TX; and Department of Neurology (O.S.), Klinikum rechts der Isar, Technische Universität München, Germany.
Petra D CravensDepartment of Neurology and Neurotherapeutics (R.Z.H., W.A.M.-L., R.D., P.D.C., O.S.), University of Texas Southwestern Medical Center, Dallas; Department of Biostatistics (G.C.), University of Alabama at Birmingham; The Moody Foundation Flow Cytometry Facility (N.L.), Children's Research Institute, University of Texas Southwestern Medical Center, Dallas; Neurology Section (O.S.), VA North Texas Health Care System, Medical Service, Dallas, TX; and Department of Neurology (O.S.), Klinikum rechts der Isar, Technische Universität München, Germany.
Olaf StüveDepartment of Neurology and Neurotherapeutics (R.Z.H., W.A.M.-L., R.D., P.D.C., O.S.), University of Texas Southwestern Medical Center, Dallas; Department of Biostatistics (G.C.), University of Alabama at Birmingham; The Moody Foundation Flow Cytometry Facility (N.L.), Children's Research Institute, University of Texas Southwestern Medical Center, Dallas; Neurology Section (O.S.), VA North Texas Health Care System, Medical Service, Dallas, TX; and Department of Neurology (O.S.), Klinikum rechts der Isar, Technische Universität München, Germany.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

objectiveTo determine the capacity, effectiveness, efficiency, and reliability of select tissue dissociation methods to isolate mononuclear cells from the CNS of mice with experimental autoimmune encephalomyelitis (EAE).

methodsAs part of an assay qualification, we tested the isolation method Percoll PLUS vs a commercially available enzymatic Neural Tissue Dissociation Kit (Kit), and the enzymes accutase and papain in C57BL/6 mice with active EAE. In a stepwise approach, we applied the following 4 criteria to each dissociation method: (1) mononuclear cell viability post-processing was required to be ≥80% per brain or spinal cord sample, (2) absolute live mononuclear cell numbers was required to be ≥5 × 10

resultsEnzymatic dissociations allowed for greatly increased cell yield and specifically allowed for downstream assays from individual brains and spinal cords in C57BL/6 mice with EAE. All enzymatic dissociations provided a more efficient and effective method for isolating mononuclear cells from brains and spinal cord. Only the Kit assay provided a significant correlation between absolute mononuclear cell numbers in the spinal cord and EAE disease severity.

conclusionsEnzymatic dissociation of CNS tissue of C57BL/6 mice with active EAE with the Kit should be the standard method. The identification of optimized CNS dissociation methods in EAE has the potential to identify cellular events that are pertinent to MS pathogenesis.

Identifiers

PMID29359175
PMCPMC5773844

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.