Comparative Proteomic Analysis Identifies EphA2 as a Specific Cell Surface Marker for Wharton's Jelly-Derived Mesenchymal Stem Cells

• Published in: International journal of molecular sciences Vol. 21; no. 17; p. 6437
• Main Authors: Al Madhoun, Ashraf, Marafie, Sulaiman K, Haddad, Dania, Melhem, Motasem, Abu-Farha, Mohamed, Ali, Hamad, Sindhu, Sardar, Atari, Maher, Al-Mulla, Fahd
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 03-09-2020; MDPI
• Subjects: adult skin fibroblasts; Biological activity; Biomarkers; Biosynthesis; Cell Differentiation; Cell Proliferation; Cell surface; Cells, Cultured; Depletion; EphA2 protein; Ephrin-A2 - metabolism; Fibroblasts; Fibroblasts - cytology; Fibroblasts - metabolism; Gene expression; Humans; Immunofluorescence; Immunological tolerance; Mass spectrometry; Mass spectroscopy; Mesenchymal stem cells; Mesenchymal Stem Cells - cytology; Mesenchymal Stem Cells - metabolism; neonate foreskin fibroblasts; Neonates; Polymerase chain reaction; Proteins; Proteome - analysis; Proteome - metabolism; proteomic analysis; Proteomics; Reverse transcription; Scientific imaging; Skin - cytology; Skin - metabolism; Stem cells; Superoxide dismutase; Surface markers; Western blotting; Wharton Jelly - cytology; Wharton Jelly - metabolism; Wharton’s jelly-derived mesenchymal stem cells; neonate foreskin fibroblasts; mass spectrometry; Wharton’s jelly-derived mesenchymal stem cells; adult skin fibroblasts; proteomic analysis
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21176437

Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) are a valuable tool in stem cell research due to their high proliferation rate, multi-lineage differentiation potential, and immunotolerance properties. However, fibroblast impurity during WJ-MSCs isolation is unavoidable because of morphological similarities and shared surface markers. Here, a proteomic approach was employed to identify specific proteins differentially expressed by WJ-MSCs in comparison to those by neonatal foreskin and adult skin fibroblasts (NFFs and ASFs, respectively). Mass spectrometry analysis identified 454 proteins with a transmembrane domain. These proteins were then compared across the different cell-lines and categorized based on their cellular localizations, biological processes, and molecular functions. The expression patterns of a selected set of proteins were further confirmed by quantitative reverse transcription polymerase chain reaction (qRT-PCR), Western blotting, and immunofluorescence assays. As anticipated, most of the studied proteins had common expression patterns. However, EphA2, SLC25A4, and SOD2 were predominantly expressed by WJ-MSCs, while CDH2 and Talin2 were specific to NFFs and ASFs, respectively. Here, EphA2 was established as a potential surface-specific marker to distinguish WJ-MSCs from fibroblasts and for prospective use to prepare pure primary cultures of WJ-MSCs. Additionally, CDH2 could be used for a negative-selection isolation/depletion method to remove neonatal fibroblasts contaminating preparations of WJ-MSCs.