Numerical measurement of viable and nonviable adipocytes and other cellular components in aspirated fat tissue
Numerical measurement of viable and nonviable adipocytes and other cellular components in aspirated fat tissue
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DOI：
10.1097/prs.0b013e31817742ed
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发表时间：
2008-07-01
影响因子：
3.6
通讯作者：
Yoshimura, Kotaro
中科院分区：
文献类型：
--
作者：
Suga, Hirotaka;Matsumoto, Daisuke;Yoshimura, Kotaro
背景资料：一个可靠的方法来测定脂肪组织中脂肪细胞和其他细胞成分的活力和数量仍有待建立。作者评估了从1g抽吸的脂肪组织和相应层获得的细胞活力和数量消化和离心之前和之后（顶层、中层和底层），使用Hoechst 33342和碘化丙啶的细胞染色和2，3-双（2-甲氧基-4-体外-5-磺基苯基）-5-[（苯基-氨基）羰基]-2H-四唑氢氧化物（XTT）和甘油-3-磷酸脱氢酶测定（n = 10）。还检查了制备的细胞（脂肪细胞、脂肪基质细胞和白色血细胞）数量与XTT和甘油-3-磷酸脱氢酶测定所得值之间的相关性（n = 5）。结果：Hoechst 33342和碘化丙啶染色法可区分脂肪细胞与脂滴、死亡脂肪细胞和脂肪细胞以外的细胞。作者从1克抽吸的脂肪组织中获得了6.9 X 10（5）个未破裂的脂肪细胞; 30%的原始脂肪细胞似乎已经破裂。XTT和甘油-3-磷酸脱氢酶测定均提供了活脂肪细胞数量与所得值之间的良好相关性，但仅甘油-3-磷酸脱氢酶测定对脂肪细胞具有严格特异性。脂肪基质细胞的脂肪细胞的比例被发现是远远大于先前described.Conclusion：单次使用或组合的活力测定在这项研究中使用的可以适当地确定脂肪细胞和其他细胞的数量，虽然它仍然很难直接评估原始细胞没有组织解离。
Background: A reliable method with which to assay viability and number of adipocytes and other cellular components in adipose tissue remains to be established.Methods: The authors assessed cell viability and number obtained from 1 g of suctioned adipose tissue and respective layers (the top, middle, and bottom layers) before and after digestion and centrifugation, using cell staining with Hoechst 33342 and propidium iodide and the 2,3-bis(2-methoxy-4-vitro-5-sulfophenyl)-5-[(phenyl-amino)carbonyl]-2H-tetrazoliumhydroxide (XTT) and glycerol-3-phosphate dehydrogenase assays (n = 10). The correlation between the number of prepared cells (adipocytes, adipose stromal cells, and white blood cells) and the resulting values from the XTT and glycerol-3-phosphate dehydrogenase assays was also examined (n = 5). The cell composition of the stromal vascular fraction isolated from the same adipose tissue was determined by multicolor flow cytometry (n = 5).Results: Hoechst 33342 and propidium iodide staining allowed distinguishing of viable adipocytes from lipid droplets, dead adipocytes, and cells other than adipocytes. The authors obtained 6.9 X 10(5) nonruptured adipocytes from 1 g of suctioned adipose tissue; 30 percent of the original adipocytes appeared to have been ruptured. Both the XTT and glycerol-3-phosphate dehydrogenase assays provided good correlations between the number of viable adipocytes and resulting values, but only the glycerol-3-phosphate dehydrogenase assay was strictly specific for adipocytes. The ratio of adipose stromal cells to adipocytes was found to be much larger than previously described.Conclusion: Single use or a combination of the viability assays used in this study can appropriately determine the number of adipocytes and other cells, although it remains difficult to assess original cells directly without tissue dissociation.