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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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Isolation, Culture, and Characterization of Primary Dermal Fibroblasts from Human Keloid Tissue
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Multifaceted array-based keloidal gene expression profiling reveals specific MDFI upregulation in keloid lesions.

M Asai1, Y Koike1, Y Kuwatsuka1

  • 1Dermatology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.

Clinical and Experimental Dermatology
|April 26, 2021
PubMed
Summary

This study identified MDFI as a key molecule in keloid development, commonly expressed in both keloid tissues and fibroblasts. MDFI-positive cells in keloid-associated lymphoid tissue may be crucial for keloid pathogenesis.

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Area of Science:

  • Dermatology
  • Molecular Biology
  • Genomics

Background:

  • Keloid lesions exhibit mesenchymal cell proliferation and excessive extracellular matrix deposition.
  • The precise molecular pathology of keloid development remains unclear despite previous studies.

Purpose of the Study:

  • To identify essential molecules involved in keloid development using microarray analysis.
  • To explore the molecular differences between keloid tissues and lesional fibroblasts.

Main Methods:

  • Microarray analysis of keloid and nonlesional skin tissues (in vivo and in vitro).
  • Gene expression comparison between tissues and cells.
  • Quantitative reverse transcription (qRT)-PCR and immunohistochemical staining for validation.

Main Results:

  • Several molecules, including PTPRD and NTM, were upregulated in keloid tissues and fibroblasts.
  • MDFI and ITGA4 were central in keloid gene coexpression networks.
  • MDFI expression was significantly elevated in keloid tissues, with MDFI-positive fibroblasts located in the keloid-associated lymphoid tissue (KALT).

Conclusions:

  • Gene expression profiles can differentiate keloid tissue from cultured fibroblasts.
  • MDFI is commonly expressed in both keloid tissues and cells.
  • MDFI-positive cells within the KALT may play a significant role in keloid pathogenesis and are relevant for in vitro studies.