title: Nonadherent culture method downregulates stem cell antigen-1 expression in mouse bone marrow mesenchymal stem cells.
abstract: Mesenchymal stem cells (MSCs) are primarily isolated by their adherence to plastic and theirin vitrogrowth 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 CD45-CD31-CD44-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.
authors.affiliation: Department of Cardiovascular Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510120, P.R. China.
authors.forename: Baoping
authors.initials: B
authors.lastname: Deng
authors.affiliation: Department of Gastroenterology, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China.
authors.forename: Weiping
authors.initials: W
authors.lastname: Deng
authors.affiliation: Center for Hematology and Regenerative Medicine (HERM), Karolinska Institute, Stockholm SE-141 86, Sweden.
authors.forename: Pingnan
authors.initials: P
authors.lastname: Xiao
authors.affiliation: Department of Cardiovascular Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510120, P.R. China.
authors.forename: Kuan
authors.initials: K
authors.lastname: Zeng
authors.affiliation: Department of Nuclear Medicine, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510120, P.R. China.
authors.forename: Shining
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Department of Transfer Medicine, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510080, P.R. China.
authors.forename: Hongwu
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Transfer Medicine, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510080, P.R. China.
authors.forename: David Yb
authors.initials: DY
authors.lastname: Deng
authors.affiliation: Department of Cardiovascular Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510120, P.R. China.
authors.forename: Yanqi
authors.initials: Y
authors.lastname: Yang
keywords: adherent culture; mesenchymal stem cells; nonadherent culture; phenotype; stem cell antigen-1