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Murine Dermal Fibroblast Isolation by FACS
Published on: January 7, 2016
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Proteomic Analysis of Human Dermal Fibroblast Conditioned Medium (DFCM)
Manira Maarof1, Yogeswaran Lokanathan1, Hj Idrus Ruszymah2
1Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaccob Latiff, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia.
The Protein Journal
|October 22, 2018
Summary
Human dermal fibroblast conditioned media (DFCM) contains proteins crucial for wound healing. Different culture media yield varying protein profiles, impacting fibroblast function and therapeutic potential.
Area of Science:
- Regenerative Medicine
- Cell Biology
- Proteomics
Background:
- Growth factors and extracellular matrix (ECM) proteins are vital for wound healing.
- Human dermal fibroblasts secrete wound-healing mediators in conditioned media (DFCM).
- DFCM composition varies with culture conditions, affecting its therapeutic efficacy.
Purpose of the Study:
- To compare the proteomic profiles of DFCM derived from human skin fibroblasts cultured in different serum-free media.
- To identify key proteins involved in wound healing processes within DFCM.
- To assess the impact of culture conditions on DFCM composition and potential wound-healing effects.
Main Methods:
- Cultured human skin fibroblasts in three serum-free media: EpiLife™ Medium (KM1), defined keratinocyte serum-free medium (KM2), and fibroblast-specific medium (FM).
- Analyzed DFCM protein content and profiles using bicinchoninic acid assay (BCA), 1D SDS-PAGE, ELISA, MALDI-TOF/TOF MS/MS, and LC-MS/MS.
- Utilized bioinformatics tools (Gene Ontology, STRING®) for protein function and pathway analysis.
Main Results:
- DFCM-KM1 and DFCM-KM2 showed higher protein concentrations than DFCM-FM.
- Proteomic analysis identified key proteins including fibronectin, serotransferrin, serpin, and serum albumin.
- Bioinformatics analysis revealed that secreted proteins are significantly associated with cellular processes, metabolic processes, growth, biological regulation, wound healing, and ECM organization.
Conclusions:
- DFCM contains proteins essential for cell adhesion, attachment, proliferation, and migration.
- The study identified specific proteins and biological processes influenced by culture conditions.
- DFCM exhibits potential for wound healing applications, with protein composition being a critical factor.
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