Deng Baoping, Deng Weiping, Xiao Pingnan, Zeng Kuan, Zhang Shining, Zhang Hongwu, Deng David Yb, Yang Yanqi
Department of Cardiovascular Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510120, P.R. China.
Department of Gastroenterology, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China.
Exp Ther Med. 2015 Jul;10(1):31-36. doi: 10.3892/etm.2015.2457. Epub 2015 Apr 29.
Mesenchymal stem cells (MSCs) are primarily isolated by their adherence to plastic and their growth characteristics. Expansion of these cells from an adherent culture is the only method to obtain a sufficient number of cells for use in clinical practice and research. However, little is known with regard to the effect of adherence to plastic on the phenotype of the cells. In the present study, bone marrow CD45CD31CD44 stem cell antigen (Sca)-1 MSCs were sorted by flow cytometry and expanded in adherent cultures. The expression levels of the adhesion molecule, Sca-1, in the adherent cultures were compared with those from nonadherent cultures at different time points. The flow cytometry results indicated that the expression levels of Sca-1 decreased in the MSCs in the nonadherent cultures grown in ultra-low-adherent plates. Furthermore, the result was confirmed by quantitative polymerase chain reaction at the same time points. Therefore, the results demonstrated that the loss of plastic adherence downregulated the expression of Sca-1. The observations may provide novel insights into the molecular mechanisms underlying plastic adherent culture.
间充质干细胞(MSCs)主要通过其对塑料的贴壁能力及其生长特性进行分离。从贴壁培养物中扩增这些细胞是获得足够数量的细胞以用于临床实践和研究的唯一方法。然而,关于贴壁于塑料对细胞表型的影响知之甚少。在本研究中,通过流式细胞术对骨髓CD45⁻CD31⁻CD44⁺干细胞抗原(Sca)-1⁺ MSCs进行分选,并在贴壁培养物中扩增。在不同时间点,将贴壁培养物中黏附分子Sca-1的表达水平与非贴壁培养物中的表达水平进行比较。流式细胞术结果表明,在超低贴壁平板中生长的非贴壁培养物中的MSCs中,Sca-1的表达水平降低。此外,在相同时间点通过定量聚合酶链反应证实了该结果。因此,结果表明失去塑料贴壁会下调Sca-1的表达。这些观察结果可能为塑料贴壁培养的分子机制提供新的见解。