Data · dataset · 2026
Establishment of a Simultaneous Single-Cell Isolation Protocol for Rat Intestinal Tissues and Analysis of Immune Cell Characteristics
Listed in ScienceDB
Description
Objective: To establish a standardized protocol for the synchronous isolation of high-viability single cells from multiple rat tissues, facilitating flow cytometric analysis of immune cell subsets and functional states across the intestinal tract and associated tissues. Methods: Small intestine, large intestine, mesentery, abdominal fat, Peyer’s patches, and mesenteric lymph nodes were harvested from Sprague-Dawley rats.
To prepare single-cell suspensions, intestinal, mesenteric, and adipose tissues underwent sequential chemical and enzymatic digestion, while lymphoid tissues were processed via mechanical dissociation. Results: Single-cell viability exceeded 85% across all samples, with yields ranging from 0.17 to 1.56 × 10⁷ cells per tissue. The distribution of immune cells exhibited distinct tissue-specific patterns:B cells were the predominant population in the majority of analyzed tissues, including Peyer’s patches, mesenteric lymph nodes, and the large intestine.
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Conversely, the small intestine was characterized by a T-cell-dominant profile, where CD4⁺ T cells accounted for (79.53 ± 7.93)% of the total T-cell population. In vitro stimulation assays revealed that IL-10+ B cells constituted 5.55% of large intestine lymphocytes, and IFN-γ+ T cells accounted for 8.91% of small intestine lymphocytes, demonstrating that this method effectively preserves cellular physiological functions.
Compared with enzymatic automatic dissociation, manual dissociation resulted in a 2.9-fold higher cell yield from the large intestine, along with elevated proportions of IL-10+ B cells and IFN-γ+ T cells post-stimulation. Conversely, automatic dissociation yielded marginally higher cell recovery from mesenteric tissues and adipose tissue. Sex had no significant effect on cell yield, viability, or immune cell subset proportions (P>0.05).
The method is consistent and robust, making it applicable to both sexes of rats and potentially extendable to other animal models. Conclusion: This standardized, multi-tissue single-cell isolation protocol offers a stable and reproducible framework for flow cytometric analysis, providing a robust methodological foundation for investigating systemic and local immune mechanisms.
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Where it is published
- DOI doi.org/10.57760/sciencedb.j00217.12451 ↗
DOI / persistent id · from scidb cn
Catalogue records · 1
- OAI-PMH record scidb.cn/oai?verb=GetRecord&metadataPrefix=oai_dc&identifier=10.57760%2… ↗
metadata API · from scidb cn
Topics
- From keywords
- Earth & Environmental Science · Engineering · Humanities · Life Sciences · Social Science
- Inferred from text
- Zoology 71%
Provenance · 1 source records, 11 field assertions
| Source | Key | Last seen | Raw |
|---|---|---|---|
| ScienceDB | 10.57760/sciencedb.j00217.12451 | 10 d ago | JSON v1 |
| Field | Assertion | Extractor | Evidence |
|---|---|---|---|
| access_level | source · scidb cn | connector:scidb_cn@1.0.0 | |
| concepts[field].anzsrc:group:3109 | enrichment · scidb cn | taxonomy-embedding@1.0.0 | title+keywords+description (71%) |
| concepts[field].local:field:earth-environmental | mapping · scidb cn | connector:scidb_cn@1.0.0 | |
| concepts[field].local:field:engineering | mapping · scidb cn | connector:scidb_cn@1.0.0 | |
| concepts[field].local:field:humanities | mapping · scidb cn | connector:scidb_cn@1.0.0 | |
| concepts[field].local:field:life-sciences | mapping · scidb cn | connector:scidb_cn@1.0.0 | |
| concepts[field].local:field:social-science | mapping · scidb cn | connector:scidb_cn@1.0.0 | |
| description | source · scidb cn | connector:scidb_cn@1.0.0 | /metadata/dc/description |
| license | source · scidb cn | connector:scidb_cn@1.0.0 | /metadata/dc/rights |
| publication_date | source · scidb cn | connector:scidb_cn@1.0.0 | |
| title | source · scidb cn | connector:scidb_cn@1.0.0 | /metadata/dc/title |