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ALCAM (CD166) as a gene expression marker for human mesenchymal stromal cell characterisation
Bas Brinkhof1, Bo Zhang1, Zhanfeng Cui1
1Institute of Biomedical Engineering, Department of Engineering Science, University of Oxford, Oxford, United Kingdom.
Gene: X
|June 19, 2020
Summary
Researchers identified new biomarkers to accurately identify mesenchymal stromal cells (MSCs) and differentiate them from fibroblasts. Activated-Leukocyte Cell Adhesion Molecule (ALCAM) showed the most promise for improving MSC characterization.
Area of Science:
- Cell Biology
- Biomarker Discovery
Background:
- Human mesenchymal stromal cells (MSCs) share surface markers (CD73, CD90, CD105) with fibroblasts, complicating identification.
- Fibroblasts are common contaminants in MSC cultures, hindering research and therapeutic applications.
- No unique MSC-specific marker has been definitively identified to date.
Purpose of the Study:
- To identify novel gene expression markers for accurate mesenchymal stromal cell (MSC) characterization.
- To differentiate MSCs from contaminating fibroblasts using molecular markers.
- To evaluate the potential of quantitative PCR (qPCR) for MSC identification.
Main Methods:
- Cultured bone marrow and umbilical cord-derived MSCs under various conditions (oxygen levels, 2D/3D culture).
- Performed quantitative PCR (qPCR) analysis on 18 candidate MSC marker genes.
- Compared gene expression levels between MSCs and fibroblasts to assess marker specificity.
Main Results:
- Standard International Society for Cell and Gene Therapy (ISCT) markers failed to distinguish MSCs from fibroblasts.
- Six genes, including ALCAM, CLIC1, EDIL3, EPHA2, NECTIN2, and TMEM47, showed potential as MSC biomarkers.
- Activated-Leukocyte Cell Adhesion Molecule (ALCAM) demonstrated high expression levels and specificity in MSCs.
Conclusions:
- Activated-Leukocyte Cell Adhesion Molecule (ALCAM) is a promising biomarker for accurate MSC identification.
- ALCAM offers improved reliability and specificity compared to existing MSC markers.
- The findings contribute to more precise characterization of MSCs for research and clinical use.
Keywords:
(q)PCR, (quantitative) polymerase chain reactionAD, adiposeAF, Amniotic FluidALCAM, Activated-Leukocyte Cell Adhesion MoleculeActivated-leukocyte cell adhesion moleculeBM, bone marrowBSG, BasiginBiomarkerCD, cluster of differentiationCLIC1, chloride intracellular channel 1CLIC4, chloride intracellular channel 4Cq, Quantification cycleDF, Dermal FibroblastsDP, Dental PulpEDIL3, EGF like repeats and discoidin domains 3ENG, EndoglinEPHA2, EPH receptor A2ER, Endoplasmatic ReticulumFACS, Fluorescence Assisted Cell SortingFN1, Fibronectin 1IGFBP7, insulin like growth factor binding protein 7ISCT, International Society for Cell and Gene TherapyITGA1, integrin subunit alpha 1LAMP1, lysosomal associated membrane protein 1LRRC59, leucine rich repeat containing 59MCAM, melanoma cell adhesion moleculeMM, Multiple MyelomaMPC, Mesenchymal Progenitor CellMSCMSC, Mesenchymal Stromal CellsNECTIN2, nectin cell adhesion molecule 2NK, Natural KillerNT5E, 5′-nucleotidase ectoOS, OsteosarcomaPL, PlacentaPPIA, peptidylprolyl isomerase APUM1, pumilio RNA binding family member 1RM, Regenerative MedicineRNARNA-seq, RNA sequencingRT, Reverse TranscriptaseRegenerative medicineSEM, Standard Error of the MeanTBP, TATA-box binding proteinTCF, Tissue Culture PlateTE, Tissue EngineeringTFRC, transferrin receptorTHY1, Thy-1 cell surface antigenTLN1, Talin 1TMEM47, transmembrane protein 47UC, umbilical cordYWHAZ, tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zetacDNA, DNA complementary to RNAqPCR
