Work

Protocol for the isolation of human buccal cells for single-cell applications

Year: 2026

Type: article

Abstract: Human buccal mucosa contains epithelial, stromal, and resident immune cell populations, but isolating these cells as a high-viability single-cell suspension is technically challenging. Here, we describe a gentle dissociation workflow for human buccal punch biopsies that combines controlled enzymatic digestion (collagenase and nuclease) with optimized speed, temperature, and incubation time to efficiently release heterogeneous cell populations while minimizing cell damage. The protocol reliably yields a heterogeneous single-cell suspension compatible with flow cytometry and single-cell RNA sequencing, with typical viabilities ≥80%.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2026.104601

Authors: Supranee Buranapraditkun, Pongsawat Rodsaward, Natcha Chottawornsak, Pattarawat Thantiworasit, Jongkonnee Wongpiyabovorn, Hiroshi Chantaphakul

Institutions: Thai Red Cross Society, Chulalongkorn University, King Chulalongkorn Memorial Hospital

Language: English

Topic: Single-cell and spatial transcriptomics

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,550