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Endotoxin contamination in isolation of lamina propria mononuclear cells
K Fukushima1, I Sasaki, K Takahashi
1The First Department of Surgery, Tohoku University School of Medicine, Sendai, Japan.
Abstract:
Because the beginning of extraction of lamina propria mononuclear cells is to obtain mucosal tissues that are exposed to luminal bacteria, the contaminated endotoxin in this step and/or the enzymes for mucosal digestion may activate mucosal macrophages and other cells. To address this issue, endotoxin levels in isolation solutions were evaluated during the extraction of lamina propria mononuclear cells from 8 control, 7 Crohn's disease and 8 ulcerative colitis specimens. Endotoxin levels were measured using Toxicolor system based on the limulus tests. Endotoxin levels were consistently below 500 pg/ml, and more importantly, these in enzyme digestion solutions were comparable among control, Crohn's disease, and ulcerative colitis. Therefore, comparative experiments using lamina propria mononuclear cells from these mucosae can be appropriately carried out, at least as far as in a comparable amount of contaminated endotoxin. However, careful consideration is required for the comparative and functional study using peripheral blood and lamina propria mononuclear cells.
Insights
Endotoxin contamination during lamina propria mononuclear cell extraction is minimal and comparable across control, Crohn's disease, and ulcerative colitis samples. This allows for reliable comparative studies of these immune cells from different patient groups.
Area of Science:
- Immunology
- Gastroenterology
Background:
- Extraction of lamina propria mononuclear cells (LPMCs) involves mucosal tissues exposed to bacteria.
- Potential endotoxin contamination and enzyme digestion may activate mucosal macrophages and other cells.
- Assessing endotoxin levels is crucial for accurate comparative studies of LPMCs.
Purpose of the Study:
- To evaluate endotoxin levels in solutions used for LPMC extraction.
- To determine if endotoxin levels differ between control, Crohn's disease, and ulcerative colitis specimens.
- To ascertain the validity of comparative functional studies using LPMCs from these groups.
Main Methods:
- Endotoxin levels were measured using the Toxicolor system (Limulus test).
- LPMCs were extracted from 8 control, 7 Crohn's disease, and 8 ulcerative colitis specimens.
- Endotoxin levels in isolation and enzyme digestion solutions were quantified.
Main Results:
- Endotoxin levels were consistently below 500 pg/ml across all samples.
- Endotoxin levels in enzyme digestion solutions were comparable between control, Crohn's disease, and ulcerative colitis groups.
- The study found comparable endotoxin contamination across the different specimen types.
Conclusions:
- Comparative experiments using LPMCs from control, Crohn's disease, and ulcerative colitis mucosae are feasible due to comparable endotoxin levels.
- Researchers can confidently conduct comparative studies on LPMCs, considering the low and consistent endotoxin contamination.
- Further consideration is needed for comparative functional studies involving peripheral blood and LPMCs.
