The Study Of Fe₃o₄ Nanoparticles Labeled Breast Cancer Stem Cells And The Influence To Their Biological Characteristics

Objective: Breast cancer tissue of MMTV-ErbB-2 transgenic mice is adopted to get its breast cancer stem cells through isolation, cultivation and identification. As the molecular imaging marker of cell in vivo tracing, Fe₃O₄ nanoparticles is used to identify the biological characteristics of BCSCs.Methods: Through enzyme digestion and stick wall to get breast cancer cells (BrCCs) from breast cancer tissue of MMTV-ErbB-2 transgenic mice, using flow cytometric(FCM) to detect the expression of CD44,CD24 (membrane surface markers) and select the CD44⁺CD24^-/low of breast cancer stem cell. The various concentration of Fe₃O₄ nanoparticles with 5mg/L,10 mg/L, 20 mg/L, 40mg/L and 80 mg/L to label the BCSCs in logarithm growing period.Through Prussian blue staining is used to detect the rate of labeling for cells in different groups, MTT method is applied to detect the cell viability so as to discuss the safe concentration of Fe₃O₄ nanoparticles, access the survival after transplation in vivo and to explore the biological characteristics of BCSCs.Results: Uniform breast cancer cells of MMTV-ErbB-2 transgenic mice can be go through primary culture and passage, Through FCM checking there are 2.2%±0.2% CD44⁺CD24^-/low cell in all the primary cells, and after culture the number of cells rise and remain stable at 80%±2%. The MTT method test showed that cell viability of Fe₃O₄ nanoparticles with 5mg/L, 10mg/L and 20mg/L teams is more than 80%, while 40mg/L and 80mg/L teams cellular viability is less than 80%; Prussian blue staining shows the labelling rate is about 20%, 90% and 100% when it is 10mg/L, 20mg/L and 40mg/L; BCSCs labeled with Fe₃O₄ nanoparticles in safety concentration, after culture Fe₃O₄ nanoparticles can pass in offspring cells; BCSCs can form new tumor after transplantation,which is proved to be breast cancer through HE staining; Prussian blue reactions where exist blue particles can be seen in breast cancer tissues via Prussian blue staining.Conclusions: BCSCs which can express CD44⁺CD24^-/low is able to be extracted successfully from the breast cencer tissue of MMTV-ErbB-2 transgenic mice The Fe₃O₄ nanoparticles concentration of 20mg/L is the most efficient and safest one to label BCSCs,and it can form new tumor after transplantation.