Son Daegu, Oh Jaehoon, Choi Taehyun, Kim Junhyung, Han Kihwan, Ha Seongyun, Lee Kyungho
Department of Plastic and Reconstructive Surgery, Keimyung University Dongsan Medical Center, Daegu, South Korea.
Ann Plast Surg. 2010 Sep;65(3):354-60. doi: 10.1097/SAP.0b013e3181bb49b8.
The purpose of this study was to determine the late decline in viability of fat cells over time for fat tissue stored at -15 degrees C and -70 degrees C after harvest from abdominal liposuction. A total of 16 females were recruited for this study. The viability of fat cell specimens was measured after freezing for 1, 3, 7, 14, 28, and 56 days. A number of viable mature adipocytes were evaluated by fluorescence microscopy after staining with fluorescein diacetate and propidium iodide. The glycerol-3-phosphate dehydrogenase activity was measured in lipoaspirates before digestion and the XTT reduction assay was performed. In addition, the XTT reduction assay was also performed on isolated lipocytes and preadipocytes.The viability of mature adipocytes was very low for both the -15 degrees C and -70 degrees C samples after 1 day of freezing (13.3% +/- 7.4% and 12.6% +/- 6.3%, respectively). There was no statistically significant difference between the samples stored at the 2 temperatures. The GPDH activity of the lipoaspirates frozen, for 1 day, at -15 degrees C and -70 degrees C was 25.1% +/- 10% and 28.7% +/- 11%, respectively. For the XTT test, the fractional enzyme activity of the lipoaspirates frozen, for 1 day, at -15 degrees C and -70 degrees C was 30.0% +/- 10.9% and 36.1% +/- 12.3%, respectively. In addition, the adipocytes had low activity from day one: 15.4% +/- 7.2% at -15 degrees C and 11.5% +/- 5.6% at -70 degrees C. Furthermore, the preadipocytes had a low activity of 8.0% +/- 6.0% at -15 degrees C and 8.6% +/- 3.8% at -70 degrees C. At 8 weeks, there were few viable mature adipocytes and the activity of the cells was very low by XTT and GPDH testing.The results of this study showed that the viability of adipocytes declined rapidly after frozen storage for 1 day at both -15 degrees C and -70 degrees C, and decreased gradually in storage after 8 weeks; at which time only approximately 5% of the fat cells were alive. These findings suggest that the present fat preservation storage techniques using a -15 degrees C freezer or a -70 degrees C deep freezer are both inadequate to maintain the viability of fat cells.
本研究的目的是确定腹部抽脂术后采集的脂肪组织在-15℃和-70℃储存时，脂肪细胞活力随时间的后期下降情况。本研究共招募了16名女性。在冷冻1、3、7、14、28和56天后测量脂肪细胞标本的活力。用荧光素二乙酸酯和碘化丙啶染色后，通过荧光显微镜评估存活的成熟脂肪细胞数量。在消化前测量抽脂物中的甘油-3-磷酸脱氢酶活性，并进行XTT还原试验。此外，还对分离的脂肪细胞和前脂肪细胞进行了XTT还原试验。冷冻1天后，-15℃和-70℃样本中成熟脂肪细胞的活力都非常低（分别为13.3%±7.4%和12.6%±6.3%）。在这两个温度下储存的样本之间没有统计学上的显著差异。在-15℃和-70℃冷冻1天的抽脂物的GPDH活性分别为25.1%±10%和28.7%±11%。对于XTT试验，在-15℃和-70℃冷冻1天的抽脂物的部分酶活性分别为30.0%±10.9%和36.1%±12.3%。此外，从第一天起脂肪细胞的活性就很低：-15℃时为15.4%±7.2%，-70℃时为11.5%±5.6%。此外，前脂肪细胞在-15℃时的活性较低，为8.0%±6.0%，在-70℃时为8.6%±3.8%。在8周时，存活的成熟脂肪细胞很少，通过XTT和GPDH检测发现细胞活性非常低。本研究结果表明，脂肪细胞在-15℃和-70℃冷冻储存1天后活力迅速下降，在储存8周后逐渐降低；此时只有约5%的脂肪细胞存活。这些发现表明，目前使用-15℃冰箱或-70℃深低温冰箱的脂肪保存储存技术都不足以维持脂肪细胞的活力。
