Source Paper
Krisztián Németh, Asada Leelahavanichkul, Peter S T Yuen, Balázs Mayer, Alissa Parmelee et al.
Nature Medicine • 2008
Objective: To evaluate the effect of bone marrow stromal cells (BMSCs) on mortality and organ function in mice with sepsis induced by cecal ligation and puncture (CLP)
This is a Cecal Ligation and Puncture (CLP) Sepsis Model protocol using mouse as the model organism. The procedure involves 9 procedural steps, 5 materials. Extracted from a 2008 paper published in Nature Medicine.
Model and subjects
mouse • Not specified in provided text • unknown • Not specified in provided text • Not specified in provided text
Study window
Estimated timing pending
Core workflow
Sepsis induction by cecal ligation and puncture (CLP) • BMSC administration • Macrophage depletion (experimental group)
Primary readouts
Key equipment and reagents
Verified items
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Induce sepsis in mice using the cecal ligation and puncture procedure
Note: This is the primary surgical intervention to create the sepsis model
“inducing sepsis by cecal ligation and puncture reduced mortality and improved organ function”
Administer bone marrow stromal cells to mice before or shortly after CLP-induced sepsis
Note: Timing of administration is flexible (before or shortly after CLP)
“Administering bone marrow stromal cells (BMSCs—also known as mesenchymal stem cells) to mice before or shortly after inducing sepsis”
Deplete macrophages in some mice to test the role of macrophages in BMSC-mediated protection
Note: This is a control/mechanistic experiment to determine if macrophages are necessary for BMSC benefit
“The beneficial effect of BMSCs was eliminated by macrophage depletion”
Pretreat mice with antibodies specific for IL-10 or IL-10 receptor to block these signaling pathways
Note: This is a control/mechanistic experiment to determine if IL-10 signaling is necessary for BMSC benefit
“pretreatment with antibodies specific for interleukin-10 (IL-10) or IL-10 receptor”
Prepare monocytes and/or macrophages from lungs of septic mice for analysis of IL-10 production
Note: Samples collected from both BMSC-treated and untreated septic mice for comparison
“Monocytes and/or macrophages from septic lungs made more IL-10 when prepared from mice treated with BMSCs versus untreated mice”
Culture LPS-stimulated macrophages with or without BMSCs to measure IL-10 production
Note: This is an ex vivo assay to test the direct effect of BMSCs on macrophage IL-10 production
“LPS-stimulated macrophages produced more IL-10 when cultured with BMSCs”
Use BMSCs lacking specific genes (Toll-like receptor 4, myeloid differentiation primary response gene-88, TNF receptor-1a, or cyclooxygenase-2) in culture experiments to determine which genes are necessary for the IL-10 production effect
Note: This mechanistic experiment identifies the molecular pathways required for BMSC-mediated macrophage reprogramming
“this effect was eliminated if the BMSCs lacked the genes encoding Toll-like receptor 4, myeloid differentiation primary response gene-88, tumor necrosis factor (TNF) receptor-1a or cyclooxygenase-2”
Measure tissue peroxidase as a marker of organ injury
Note: Used to assess organ function following sepsis and BMSC treatment
“K.N. formulated the molecular mechanism hypothesis and designed and performed in vitro and ex vivo assays; B.M. performed the measurements for tissue peroxidase”
Perform flow cytometry analysis to characterize cell populations
Note: Used for detailed cellular analysis
“I.J. performed FACS experiments”
This section explains what the experiment is doing, which readouts matter, what the data artifacts usually look like, and how the analysis should flow from raw capture to reported result.
To evaluate the effect of bone marrow stromal cells (BMSCs) on mortality and organ function in mice with sepsis induced by cecal ligation and puncture (CLP)
Objective
To evaluate the effect of bone marrow stromal cells (BMSCs) on mortality and organ function in mice with sepsis induced by cecal ligation and puncture (CLP)
Subjects
From papermouse • Not specified in provided text • unknown • Not specified in provided text • Not specified in provided text
Cohort notes
From paperSome mice were genetically modified to lack specific genes (Toll-like receptor 4, myeloid differentiation primary response gene-88, TNF receptor-1a, or cyclooxygenase-2)
Sepsis induction by cecal ligation and puncture (CLP) (Not specified in provided text)
BMSC administration (Before or shortly after sepsis induction)
Macrophage depletion (experimental group) (Not specified in provided text)
Pretreatment with blocking antibodies (experimental group) (Not specified in provided text)
Mortality rate in septic mice
From paperNot specified in provided text
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Organ function/injury markers
From paperNot specified in provided text
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
IL-10 production by monocytes and macrophages from septic lungs
From paperNot specified in provided text
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
IL-10 production by LPS-stimulated macrophages cultured with BMSCs
From paperNot specified in provided text
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Mortality rate in septic mice
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Organ function/injury markers
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
IL-10 production by monocytes and macrophages from septic lungs
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
IL-10 production by LPS-stimulated macrophages cultured with BMSCs
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
Preprocessing / cleaning
Not specified in provided text
Scoring or quantification
Quantify the primary readouts for this experiment: Mortality rate in septic mice; Organ function/injury markers; IL-10 production by monocytes and macrophages from septic lungs; IL-10 production by LPS-stimulated macrophages cultured with BMSCs.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Mortality rate in septic mice, Organ function/injury markers, IL-10 production by monocytes and macrophages from septic lungs, IL-10 production by LPS-stimulated macrophages cultured with BMSCs.
Source links and direct wording from the methods section for validation and deeper review.
Citation
Krisztián Németh et al. (2008). Bone marrow stromal cells attenuate sepsis via prostaglandin E2–dependent reprogramming of host macrophages to increase their interleukin-10 production. Nature Medicine
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Current status surfaces were computed from experiment data updated Feb 28, 2026.
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Jump back into the original paper or the methods evidence section when you need exact wording, exclusions, or method-specific caveats.
This protocol has structured steps plus evidence quotes, and is ready for canonical sync.
Steps
9
Evidence Quotes
14
Protocol Items
5
Linked Products
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Canonical Sync
Pending
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Computed from the current experiment record updated Feb 28, 2026.
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Steps
9
Evidence
14
Specific Products
0/0
Canonical Sync
Pending
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Computed from the current experiment record updated Feb 28, 2026.
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