ABSTRACT
Objective To analyze the compatibility of bone marrow mesenchymal stem cells (BMSCs) with amphiphilic peptide three-dimensional gel.Methods Three healthy 3-week old SD rats were taken for separating femur and tibia to obtain BMSCs,the BMSCs surface antigen was detected by flow cytometry;the 10 mg/mL RGD-cyclic amphiphilic peptide solution was added into the same volume of DMEM/F12 culture medium,after a few seconds,which was self assemble into three-dimensional gel.Three dimensional gel structure was observed by transmission electron microscope (TEM).1 × 106 cells/mL BMSCs suspension and RGDcyclic amphiphilic peptide were mixed to form a 3D culture system,1 × 106 cells/mL BMSCs suspension was mixed with polylysine to form a 2D culture system,the serum-free culture was conducted;the CCK-8 method was used to observe the cell growth situation,calcein acetoxy methyl ester/propidium iodide (PI) double standard staining was performed.The effect of RGD-cyclic amphiphilic peptide on the proliferation of BMSCs was observed by fluorescence microscopy.Results The separated and cultured BMSCs highly expressed CD29 and CD90,but lowly expressed or did not express CD34 and CD45;TEM showed that the gel was composed of multiple empty nanofibers with the nanofiber diameter of 2-5 nm and length of 100-1 000 nm;the molecular weight of synthetic peptides detected by mass spectrometry (MS) was 1 256.37,which was consistent with the theoretical value;the HPLC analysis showed that RGD-amphiphilic peptide purity was 95.88 ％;the calcein acetoxyl methyl ester/PI double staining showed that in the 3D culture system,a few BMSCs died after 30 min and the cells began to proliferate after 12 h,the proliferation was more active than that of 2D culture,and the difference was statistically significant (P＜0.05);CCK-8 cell count showed that the proliferation activity of 3D culture system was higher than that of 2D culture system,and the difference was statistically significant (P＜0.05).Conclusion RGD amphiphilic peptide has a good biocompatibility with BMSCs,and may become the tissue engineering scaffold material.