Abstract:

BACKGROUND: Dil derivative CM-Dil easily insets, disperses and stably binds to the whole cell membrane because of water-solubility and chloromethylated active sulfydryl, which leads to rapid, even and long-lasting staining.
OBJECTIVE: To further validate the feasibility of fluorescent dye CM-Dil in labeling rat adipose derived stem cells.
METHODS: Rat inguinal fat tissue was resected and adipose derived stem cells were isolated in vitro by collagenase digestion. Passage 3 cells were divided into a control group and an experimental group. The experimental group cells were labeled with 4 mg/L CM-Dil. At 6, 12, 24 and 48 hours, CM-Dil-labeled adipose derived stem cells were traced in vivo.
RESULTS AND CONCLUSION: Under fluorescent microscope, CM-Dil labeled cells showed cytoplasm and cell membrane with red fluorescence, CM-Dil labeled adipose derived stem cells exhibited a shuttle-shaped normal appearance. CM-Dil positive rate was 100%. In the early period, the cells were fluorescent ring-shaped, and 48 hours later, fluorescent granules increased and fluorescence intensity enhanced. No fluorescence was found in cell nucleus. There was no significant difference in cell morphology, content of lactic dehydrogenase in supernatant and MTT value between experimental group and control group (P > 0.05). At 4 hours after cell transplantation, cells with red fluorescence could be observed in the heart and lung tissue. CM-DiI can effectively label adipose derived stem cells cultured in vitro and express stably in cells. CM-Dil-labeled cells have good morphology and no toxicity to living cells.