A method of limited dilution to sort human osteosarcoma 143-B stem cells and its identification

Objective: To explore the methods of cell sorting and indentification of human osteosarcoma 143-B stem cells.Methods: The 143-B cells were treated by limited dilution method for stem cell sorting, and the expression levels of Nanog and OCT4 were determined. The multipotentials of osteogenic and adipogenic differentiation were detected. The ablility of tumorigenesis was examined by tumor formation experiment in vivo, and the drug resistance assay was performed to detect the effect of cisplatin on half inhibitory concentration (IC50).Results: The monoclonal 143-L cells obtained by cell sorting had a high ability of sphere formation. The Nanog and OCT4 expression levels on obtained monoclonal cells (143-L cells) were significantly higher than those in 143-B cells. Meanwhile, the 143-B cells had multipotentials of osteogenic, adipogenic and chondrogenic differentiation. Furthermore, the tumorigenicity in nude mice and drug resistant ability of 143-L cells (the IC50 values of 143-B and 143-L cells were 0.83 and 2.51 μg/mL, respectively) were stronger than those of 143-B cells.Conclusion: The limited dilution method can be used to sort 143-B stem cells. DOI:10.3781/j.issn.1000-7431.2015.11.527