Son Daegu, Choi Taehyun, Yeo Hyeonjung, Kim Junhyung, Han Kihwan
From the *Department of Plastic and Reconstructive Surgery, Keimyung University School of Medicine, Daegu; and †Department of Plastic and Reconstructive Surgery, Seoul National University College of Medicine, Seoul, South Korea.
Ann Plast Surg. 2014 May;72(5):589-93. doi: 10.1097/SAP.0b013e318268a85d.
Different researchers have recommended different lipoaspirate centrifugation speeds and times, probably due to the limits in fat cell viability assays. We assessed fat cell viability using a fluorescein diacetate and propidium iodide (FDA-PI) stain and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay after harvesting syringe liposuction and spun with different centrifugation speeds to determine the optimal conditions. Lipoaspirates, harvested from 13 donors, were transferred into a centrifuge tube and spun at 1000, 3000, and 4000 rpm for 3 minutes. Mature adipocytes and adipose stem cells were isolated and tested with a direct counting of FDA-PI-stained cells under fluorescence microscope and XTT assay. We incubated adipocytes and adipose stem cells for 1 day and 3 days, and we compared both of them with fresh samples to evaluate the influence of culturing condition on fat cell viability. Centrifugation speeds from 1000 rpm to 4000 rpm for 3 minutes showed no change in the percentage of adipocytes and adipose stem cell viability not only in the fresh samples but also in the cultured samples (1 day and 3 days). Centrifugation speeds under 4000 rpm do not change the percentage of fat cell viability. To differentiate viable cells from dying or dead mature adipocytes and oil accurately, combinations of viability tests are essential.
不同的研究人员推荐了不同的脂肪抽吸物离心速度和时间，这可能是由于脂肪细胞活力测定方法存在局限性。我们在收集注射器抽脂物后，以不同的离心速度进行离心，使用双醋酸荧光素和碘化丙啶（FDA-PI）染色以及2,3-双（2-甲氧基-4-硝基-5-磺基苯基）-2H-四唑-5-羧基苯胺（XTT）测定法来评估脂肪细胞活力，以确定最佳条件。从13名捐赠者身上收集的脂肪抽吸物被转移到离心管中，分别以1000、3000和4000转/分钟的速度离心3分钟。分离出成熟脂肪细胞和脂肪干细胞，并在荧光显微镜下直接计数FDA-PI染色的细胞，同时进行XTT测定。我们将脂肪细胞和脂肪干细胞分别培养1天和3天，并将它们与新鲜样本进行比较，以评估培养条件对脂肪细胞活力的影响。在新鲜样本以及培养样本（1天和3天）中，1000转/分钟至4000转/分钟离心3分钟的速度下，脂肪细胞和脂肪干细胞活力的百分比均未发生变化。4000转/分钟以下的离心速度不会改变脂肪细胞活力的百分比。为了准确区分活细胞与濒死或死亡的成熟脂肪细胞及油脂，活力测试的组合至关重要。
