[Comparison of the efficiency of three techniques for labeling human adipose-derived stem cells].

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

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• Level B · Emerging clinical evidence with positive signals
• Level C · Early human research exploring benefits
• Level D · Scientific groundwork from lab and animal studies
• Emerging · Emerging topic under active research

Abstract (original English)

Objective To explore the optimal methods for labeling human adipose-derived stem cells (ASCs). Methods ASCs were isolated by collagenase digestion and density gradient centrifugation, and their cell surface markers and ability to differentiate into the adipogenic, chondrogenic, and osteogenic lineages were examined in vitro. Three different cell labeling methods, namely 5 µl DiI, 10 µg/ml BrdU and 50 MOI adenovirus carrying GFP, were used for ASC labeling, and the labeling efficiency were compared at different time points and in different passages using fluorescent microscope. Results The isolated ASCs were capable of differentiating into adipogenic, osteogenic, chondrogenic lineages with positive stem cell marker expression. At 48 h after DiI staining, 100% of the ASCs emitted red fluorescence in the cytoplasm with fluorescent-negative nuclei, but the fluorescence intensity declined quickly after cell passaging. With 10 µg/ml BrdU, 90% of the cells showed green fluorescence in the cell nuclei at 48 h after the labeling, but the positivity rate also decreased gradually after cell passaging. Cell labeling with GFP adenovirus showed more stable labeling efficiency, and green fluorescence was detected at 24 h after labeling, and even till 5 days later more than 90% of the ASCs remained positive without an obvious attenuation of the fluorescent intensity even after cell passaging.

Evidence level

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

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