Wetzig Andrew, Alaiya Ayodele, Al-Alwan Monther, Pradez Christian Benedict, Pulicat Manogaran S, Al-Mazrou Amer, Shinwari Zakia, Sleiman Ghida Majed, Ghebeh Hazem, Al-Humaidan Hind, Gaafar Ameera, Kanaan Imaduddin, Adra Chaker
Stem Cell & Tissue Re-engineering Program, King Faisal Specialist Hospital and Research Centre, PO Box 3354, Riyadh 11211, Kingdom of Saudi Arabia.
BMC Cell Biol. 2013 Dec 5;14:54. doi: 10.1186/1471-2121-14-54.
Mesenchymal stem cells have properties that make them amenable to therapeutic use. However, the acceptance of mesenchymal stem cells in clinical practice requires standardized techniques for their specific isolation. To date, there are no conclusive marker (s) for the exclusive isolation of mesenchymal stem cells. Our aim was to identify markers differentially expressed between mesenchymal stem cell and non-stem cell mesenchymal cell cultures. We compared and contrasted the phenotype of tissue cultures in which mesenchymal stem cells are rich and rare. By initially assessing mesenchymal stem cell differentiation, we established that bone marrow and breast adipose cultures are rich in mesenchymal stem cells while, in our hands, foreskin fibroblast and olfactory tissue cultures contain rare mesenchymal stem cells. In particular, olfactory tissue cells represent non-stem cell mesenchymal cells. Subsequently, the phenotype of the tissue cultures were thoroughly assessed using immuno-fluorescence, flow-cytometry, proteomics, antibody arrays and qPCR.
Our analysis revealed that all tissue cultures, regardless of differentiation potential, demonstrated remarkably similar phenotypes. Importantly, it was also observed that common mesenchymal stem cell markers, and fibroblast-associated markers, do not discriminate between mesenchymal stem cell and non-stem cell mesenchymal cell cultures. Examination and comparison of the phenotypes of mesenchymal stem cell and non-stem cell mesenchymal cell cultures revealed three differentially expressed markers - CD24, CD108 and CD40.
We indicate the importance of establishing differential marker expression between mesenchymal stem cells and non-stem cell mesenchymal cells in order to determine stem cell specific markers.
间充质干细胞具有适合治疗用途的特性。然而，间充质干细胞在临床实践中的应用需要标准化的特定分离技术。迄今为止，尚无用于间充质干细胞特异性分离的确切标志物。我们的目的是鉴定间充质干细胞与非干细胞间充质细胞培养物之间差异表达的标志物。我们比较并对比了间充质干细胞丰富和稀少的组织培养物的表型。通过初步评估间充质干细胞的分化，我们确定骨髓和乳腺脂肪培养物富含间充质干细胞，而在我们的研究中，包皮成纤维细胞和嗅觉组织培养物中含有稀少的间充质干细胞。特别是，嗅觉组织细胞代表非干细胞间充质细胞。随后，使用免疫荧光、流式细胞术、蛋白质组学、抗体阵列和定量聚合酶链反应对组织培养物的表型进行了全面评估。
我们的分析表明，所有组织培养物，无论其分化潜能如何，都表现出非常相似的表型。重要的是，还观察到常见的间充质干细胞标志物和成纤维细胞相关标志物并不能区分间充质干细胞和非干细胞间充质细胞培养物。对间充质干细胞和非干细胞间充质细胞培养物的表型进行检查和比较，发现了三种差异表达的标志物——CD24、CD108和CD40。
我们指出了确定间充质干细胞和非干细胞间充质细胞之间差异标志物表达对于确定干细胞特异性标志物的重要性。
