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Updated: Jul 25, 2025

Isolation, Culture, and Characterization of Primary Dermal Fibroblasts from Human Keloid Tissue

Isolation, Culture, and Characterization of Primary Dermal Fibroblasts from Human Keloid Tissue

Published on: July 28, 2023

Single-Cell Transcriptome Analysis Identifies Subclusters with Inflammatory Fibroblast Responses in Localized

Giffin Werner¹, Anwesha Sanyal¹, Emily Mirizio¹

¹Department of Pediatrics (Rheumatology), University of Pittsburgh, Pittsburgh, PA 15224, USA.

International Journal of Molecular Sciences

|June 28, 2023

Summary

Localized scleroderma (LS) involves skin inflammation and fibrosis. Single-cell RNA sequencing identified unique fibroblast populations in LS skin, offering new insights into disease mechanisms.

Keywords:

CXCL10 CXCL9 CXCR3 ligands IL-6 cell communication fibroblasts fibrosis inflammation localized scleroderma morphea single-cell RNA sequencing skin

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Murine Dermal Fibroblast Isolation by FACS

Murine Dermal Fibroblast Isolation by FACS

Published on: January 7, 2016

Area of Science:

Dermatology
Immunology
Genomics

Background:

Localized scleroderma (LS) is an autoimmune skin disease characterized by inflammation and fibrosis, leading to collagen deposition.
Its pathophysiology is often inferred from systemic sclerosis (SSc) due to similar skin histopathology.
LS remains significantly understudied compared to SSc.

Purpose of the Study:

To investigate the cellular and molecular landscape of affected skin in localized scleroderma using single-cell RNA sequencing.
To identify distinct fibroblast subpopulations and their gene expression profiles in LS.
To uncover novel cellular players and pathways involved in LS pathogenesis.

Main Methods:

Analysis of skin biopsies from 14 patients with localized scleroderma (pediatric and adult) and 14 healthy controls.
Application of single-cell RNA sequencing (scRNA-seq) to profile individual cells.
Bioinformatic analysis to identify fibroblast subclusters, gene expression patterns, and cell-cell communication networks.

Main Results:

Identification of 12 distinct fibroblast subclusters in localized scleroderma skin.
Fibroblast populations exhibited significant inflammatory gene expression, including interferon (IFN) and HLA-associated genes.
A specific myofibroblast-like cluster (SFRP4/PRSS23) was more prevalent in LS and showed upregulation of SSc-associated genes and CXCR3 ligands (CXCL9/10/11).
A unique CXCL2/IRF1 cluster with a strong inflammatory signature, including IL-6, was identified in LS and influenced by macrophages.

Conclusions:

Single-cell RNA sequencing revealed novel fibroblast subpopulations and gene signatures in localized scleroderma.
These findings highlight distinct inflammatory pathways and cellular interactions driving fibrosis in LS.
The identified fibroblast clusters and signatures represent potential therapeutic targets for localized scleroderma.