不同冻存时间对人颗粒脂肪活性影响的实验研究  

Experimental study on the effect of different cryopreservation time on human fat granule

作　　者：魏国茜 顾洛莎 伍明政 白洁[1] 顾劲松[1] 于江[1] 闫润虎[1] 张怡[1] WEI Guoqian;GU Luosha;WU Mingzheng;BAI Jie;GU Jinsong;YU Jiang;YAN Runhu;ZHANG Yi(Cosmetic Medical College, Dalian Medical University, Liaoning Province, Dalian 116044, China;Medical College, Zhengzhou University, He′nan Province, Zhengzhou 450001, China;Department of Outpatient, Guangmei Medical Plastic and Cosmetic, Guangdong Province, Guangzhou 510507, China)

机构地区：[1]大连医科大学美容医学院,辽宁大连116044 [2]郑州大学医学院,河南郑州450001 [3]广东省广美整形美容医疗门诊部,广东广州510507

出　　处：《中国医药导报》2019年第6期16-19,F0004,共5页China Medical Herald

基　　金：校企横向合作课题(HX2014002)

摘　　要：目的探索不同冻存时间对人颗粒脂肪活力的影响,寻求更为满意的颗粒脂肪冻存时间,为临床自体脂肪移植提供理论依据。方法收集2016年6～10月广州广美整形美容医疗门诊部20～40岁行大腿内侧自体脂肪抽吸移植术女性41例的脂肪标本,经生理盐水漂洗2次静置分离后加入相应留存脂肪量等体积的低温冻存保护液于-196℃(液氮)冻存。选择冻存3个月(A组)和6个月(B组)的标本复温,通过苏木精-伊红(HE)及台盼蓝染色观察冻存脂肪细胞形态并计算脂肪细胞存活率,肌酸激酶测定酶活力、免疫组化CD105检测脂肪干细胞(ADSCs)数量,比较A、B两组脂肪细胞活力变化情况。结果脂肪细胞计数分析脂肪细胞存活率及肌酸激酶测定结果显示,在相同低温条件下两组脂肪细胞活力差异无统计学意义(P> 0.05);HE染色显微镜下比较脂肪细胞形态发现,A、B两组脂肪细胞形态基本正常,仅于A组见少数ADSCs。结论使用液氮冻存人颗粒脂肪组织安全有效,于-196℃下冻存6个月,甚至更长时间,是较理想的冻存条件。Objective To explore the effect of different cryopreservation time on human adipose cell vitality, and seek a more satisfactory granular fat cryopreservation time, to provide a theoretical reference for autologous fat transplantation in clinic. Methods The fat granule samples from interior thigh of 41 healthy women (20-40 years old) that underwent conventional liposuction were collected in Guangmei Medical Plastic and Cosmetic Outpatient Department from June to October 2016. All fat granule samples were cryopreserved in liquid nitrogen freezer of -196℃ after washed twice by physiological saline and placed quiescently to isolate them from the physiological saline and then an equal volume of freezing protection liquid was added into the fat granule samples. The samples that had been stored for 3 months (group A) and 6 months (group B) were selected to test the activity of frozen fat after thawed. The morphology and the death cell percentage of cryopreserved granule fat cells was observed and calculated respectively by hematoxylin-eosin (HE) staining and trypan-blue staining. The creatine kinase level was tested to calculate fat cell enzyme vitality. The CD105 on adipose derived stem cells (ADSCs) was detected by immunohistochemical methods to compare the effect of different cryopreservation time on the fat cell activity in vitro. Results Fat cell count analysis on the survival rate of fat cells and the determination of creatine kinase showed no statistically significant differences in the fat cell survival rates between group A and group B under the same cryopreservation temperature (P > 0.05). The comparative observation of the fat cell morphology by HE staining under the microscope showed that all cell membranes keep well in the two groups. Immunohistochemical staining of CD105 showed that there were only a few of ADSCs in group A. Conclusion Cryopreservation of human granular adipose tissue with liquid nitrogen is safety and effective. The fat cells can be cryopreserved at -196℃ for 6 months or even longer t

关 键 词：颗粒脂肪 冷冻保存 储存时间 脂肪细胞活力 

分 类 号：R622.9[医药卫生—整形外科]