Fluorescence labeling for human bone marrow mesenchymal stem cells with PKH26 / 中华检验医学杂志

ABSTRACT

Objective To establish a method of labeling human mesenchymal stem cells (MSCs) with PKH26 in vitro.Methods MSCs were cultured and labeled with PKH26 according to the manufacturer's instruction.The growth,fluorescence intensity and serial subcuhivation of labeled MSCs were analyzed with the confocal laser microscope and the flow cytometry.The biological characteristics of labeled MSCs were investigated by RT-PCR.Results The labeled MSCs appeared red fluorescence and the labeling rate was 100 percent.During serial subcuhivation of labeled MSC from passage 1 to passage 7,the fluorescence intensity and the labeling rate of MSCs were gradually decreased.The biological features such as morphology,growth,expression level of nucleostemin and GAPDH gene and capability of differentiation into osteoblast in vitro were not affected by labeling.Conclusion Labeling the human MSCs with PKH26 is an effective and practical method,which can be used as an important tool in the study on the homing, plasticity and transplantation of MSCs.