title: Derivation, characterization and gene modification of cynomolgus monkey mesenchymal stem cells.
abstract: Mesenchymal stem cells (MSCs) have received considerable attention in recent years. Particularly exciting is the prospect that MSCs could be differentiated into specialized cells of interest, which could then be used for cell therapy and tissue engineering. MSCs derived from nonhuman primates could be a powerful tool for investigating the differentiation potential in vitro and in vivo for preclinical research. The purpose of this study was to isolate cynomolgus mesenchymal stem cells (cMSCs) from adult bone marrow and characterize their growth properties and multipotency. Mononuclear cells were isolated from cynomolgus monkey bone marrow by density-gradient centrifugation, and adherent fibroblast-like cells grew well in the complete growth medium with 10 microM Tenofovir. cMSCs expressed mesenchymal markers, such as CD29, CD105, CD166 and were negative for hematopoietic markers such as CD34, CD45. Furthermore, the cells were capable of differentiating into osteogenic, chondrogenic, and adipogenic lineages under certain conditions, maintaining normal karyotype throughout extended culture. We also compared different methods (lipofection, nucleofection and lentivirus) for genetic modification of cMSCs and found lentivirus proved to be the most effective method with transduction efficiency of up to 44.6% and lowest level of cell death. The cells after transduction stably expressed green fluorescence protein (GFP) and maintained the abilities to differentiate down osteogenic and adipogenic lineages. In conclusion, these data showed that cMSCs isolated from cynomolgus bone marrow shared similar characteristics with human MSCs and might provide an attractive cell type for cell-based therapy in higher-order mammalian species disorder models.
authors.affiliation: Center for Stem Cell Biology and Tissue Engineering, The Key Laboratory for Stem Cells and Tissue Engineering, Sun Yat-Sen University, Ministry of Education, Guangzhou 510080, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Ke
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.forename: Weihua
authors.initials: W
authors.lastname: Yu
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Liu
authors.forename: Chang
authors.initials: C
authors.lastname: Liu
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.forename: Frank Fuxiang
authors.initials: FF
authors.lastname: Mao
authors.forename: Liangming
authors.initials: L
authors.lastname: Zhang
authors.forename: Xiuming
authors.initials: X
authors.lastname: Zhang
authors.forename: Bruce T
authors.initials: BT
authors.lastname: Lahn
authors.forename: Andy Peng
authors.initials: AP
authors.lastname: Xiang
mesh_terms: D000818:Animals; D001853:Bone Marrow; D002454:Cell Differentiation; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D005434:Flow Cytometry; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007621:Karyotyping; D008252:Macaca fascicularis; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D014162:Transfection
chemical_list: D049452:Green Fluorescent Proteins