不同配比低温保护剂对冻存人颗粒脂肪裸鼠移植的影响
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摘要
目的探讨不同配比低温保护剂对冻存后人颗粒脂肪裸鼠移植的影响,以优化低温保护剂的选择。方法取人大腿内侧吸脂术后多余的颗粒脂肪,分别等量存入A、B两种不同配比的低温保护剂中,分别为A组和B组,置于-80℃冻存6个月。6个月后将标本复苏,移植于裸鼠背部皮下,各移植点注射0.2 m L颗粒脂肪。4周后处死裸鼠,取出移植物,进行体积、重量检测,并观察其苏木精-伊红(HE)染色、透射电镜下的情况。结果 A组剩余脂肪平均体积大于B组,差异有统计学意义(P <0.05)。HE染色示A组新生血管多于B组;透射电镜下A组脂肪细胞更加完整,新生脂肪细胞较多。结论 A组低温保护剂长期冻存的脂肪活性较高,且可用小牛血清取代胎牛血清;脂肪组织深低温冻存半年以后依然可以进行有效移植。
Objective To investigate the effects of cryoprotective agents with different proportions on transplantation of frozen human granular fat in nude mice in order to optimize the selection of cryoprotectants. Methods The excess granular fat after liposuction on the inner side of the thigh was taken into the cryoprotective agents of A and B in different proportions, group A and group B respectively and they were stored at-80°C for 6 months. After 6 months, the specimens were resuscitated and transplanted subcutaneously into the back of nude mice, and 0.2 mL granular fat was injected at each transplant site. After 4 weeks, the nude mice were sacrificed, and the grafts were taken out for volume and weight detection, and observed under hematoxylin-eosin(HE) staining and transmission electron microscopy. Results The mean volume of residual fat in group A was greater than that in group B, and the difference was statistically significant(P < 0.05). HE staining showed that the neovascularization of group A was more than that of group B. Under transmission electron microscope, the fat cells of group A were more complete and there were more new fat cells. Conclusion The long-term frozen fat of group A cryoprotective agents has higher activity, and fetal bovine serum can be replaced by calf serum. The adipose tissue can still be effectively transplanted after cryopreservation for half a year.
Objective To investigate the effects of cryoprotective agents with different proportions on transplantation of frozen human granular fat in nude mice in order to optimize the selection of cryoprotectants. Methods The excess granular fat after liposuction on the inner side of the thigh was taken into the cryoprotective agents of A and B in different proportions, group A and group B respectively and they were stored at-80°C for 6 months. After 6 months, the specimens were resuscitated and transplanted subcutaneously into the back of nude mice, and 0.2 mL granular fat was injected at each transplant site. After 4 weeks, the nude mice were sacrificed, and the grafts were taken out for volume and weight detection, and observed under hematoxylin-eosin(HE) staining and transmission electron microscopy. Results The mean volume of residual fat in group A was greater than that in group B, and the difference was statistically significant(P < 0.05). HE staining showed that the neovascularization of group A was more than that of group B. Under transmission electron microscope, the fat cells of group A were more complete and there were more new fat cells. Conclusion The long-term frozen fat of group A cryoprotective agents has higher activity, and fetal bovine serum can be replaced by calf serum. The adipose tissue can still be effectively transplanted after cryopreservation for half a year.
引文
[1]Fournier PF.Facial recontouring with fat grafting[J].Dermatol Clin,1990,8(3):523-537.
[2]Jackson RF.Frozen fat-does it work[J].Am J Cosmet Surg,1997,14(3):339-343.
[3]Sattler G,Sommer B.Liporecycling:immediate and delayed[J].Am J Cosmet Surg,1997,14:311-316.
[4]Narins RS.Long term sterility of fat frozen for up to 24months[J].J Drugs Dermatol,2003,2(5):505-510.
[5]张广宇,于冬梅,郝立君,等.不同低温条件保存的大鼠脂肪自体移植后成活率的探究[J].中国整形美容外科,2016:27(3):181-185.
[6]Hivernaud V,Lefourn B,Robard M,et al.Autologous fat grafting:A comparative study of four current commercial protocols[J].J Plast Reconstr Aesthet Surg,2017,70(2):248-256.
[7]邓颖,刘少倩,谢红炬,等.海藻糖对脂肪细胞冻存后存活率的影响[J].中南大学学报:医学版,2017;42(5):507-510.
[8]仇侃敏,吴蒙,梁耀蝉,等.不同低温保护剂保存的颗粒性脂肪组织活力及移植成活率分析[J].基础医学与临床,2014,34(4):536-540.
[9]程爱娟,刘春霞.冻存颗粒脂肪组织移植成活率的实验研究[J].菏泽医学专科学校学报,2017,29(2):5-7,11.
[10]Bellini E,Grieco MP,Raposio E.The science behind autologous fat grafting[J].Ann Med Surg(Lond),2017,24:65-73.
[11]Simonacci F,Bertozzi N,Grieco MP,et al.Autologous fat transplantation for breast reconstruction:A literature review[J].Ann Med Surg(Lond),2016,12:94-100.
[12]Chen YW,Wang JR,Liao X,et al.Effect of suction pressures on cell yield and functionality of the adipose-derived stromal vascular fraction[J].J Plast Reconstr Aesthet Surg,2017,70(2):257-266.
[13]Simonacci F,Bertozzi N,Grieco MP,et al.Procedure,applications,and outcomes of autologous fat grafting[J].Ann Med Surg(Lond),2017,20:49-60.
[14]虞效益,张绍志,范菊莉,等.低温保护剂渗透关节软骨研究进展[J].中国生物医学工程学报,2015,34(4):481-486.
[15]Yun Y,Li ZX,Jun DJ,et al.Osmotic Injury to Porcine Oocytes during Cryoprotective Agents Addition and Removal Processes[J].Int J Refrig,2017,38(3):114-118.
[16]张浩.牛血清在细胞培养中的作用与质量要求[J].科技情报开发与经济,2001,11(3):46-47.
[17]S覬ndergaard RH,Follin B,Lund LD,et al.Senescence and quiescence in adipose-derived stromal cells:Effects of human platelet lysate,fetal bovine serum and hypoxia[J].Cytotherapy,2017,(19):95-106.
[18]林常敏,李宇,纪影畅,等.毛乳头细胞培养基血清浓度的探索[J].中国美容整形外科杂志,2008,19(3):218-220.
[19]金亦宏,马欣,周莉薇.国产牛血清的现状[J].中国生物制品学杂志,2004,l7(4):254-256.
[20]吕维敏,黄钢妹,曾叶,等.生物材料冻存中低温保护剂的作用机理及实验研究[J].低温与超导,2014,42(5):12-16.
[2]Jackson RF.Frozen fat-does it work[J].Am J Cosmet Surg,1997,14(3):339-343.
[3]Sattler G,Sommer B.Liporecycling:immediate and delayed[J].Am J Cosmet Surg,1997,14:311-316.
[4]Narins RS.Long term sterility of fat frozen for up to 24months[J].J Drugs Dermatol,2003,2(5):505-510.
[5]张广宇,于冬梅,郝立君,等.不同低温条件保存的大鼠脂肪自体移植后成活率的探究[J].中国整形美容外科,2016:27(3):181-185.
[6]Hivernaud V,Lefourn B,Robard M,et al.Autologous fat grafting:A comparative study of four current commercial protocols[J].J Plast Reconstr Aesthet Surg,2017,70(2):248-256.
[7]邓颖,刘少倩,谢红炬,等.海藻糖对脂肪细胞冻存后存活率的影响[J].中南大学学报:医学版,2017;42(5):507-510.
[8]仇侃敏,吴蒙,梁耀蝉,等.不同低温保护剂保存的颗粒性脂肪组织活力及移植成活率分析[J].基础医学与临床,2014,34(4):536-540.
[9]程爱娟,刘春霞.冻存颗粒脂肪组织移植成活率的实验研究[J].菏泽医学专科学校学报,2017,29(2):5-7,11.
[10]Bellini E,Grieco MP,Raposio E.The science behind autologous fat grafting[J].Ann Med Surg(Lond),2017,24:65-73.
[11]Simonacci F,Bertozzi N,Grieco MP,et al.Autologous fat transplantation for breast reconstruction:A literature review[J].Ann Med Surg(Lond),2016,12:94-100.
[12]Chen YW,Wang JR,Liao X,et al.Effect of suction pressures on cell yield and functionality of the adipose-derived stromal vascular fraction[J].J Plast Reconstr Aesthet Surg,2017,70(2):257-266.
[13]Simonacci F,Bertozzi N,Grieco MP,et al.Procedure,applications,and outcomes of autologous fat grafting[J].Ann Med Surg(Lond),2017,20:49-60.
[14]虞效益,张绍志,范菊莉,等.低温保护剂渗透关节软骨研究进展[J].中国生物医学工程学报,2015,34(4):481-486.
[15]Yun Y,Li ZX,Jun DJ,et al.Osmotic Injury to Porcine Oocytes during Cryoprotective Agents Addition and Removal Processes[J].Int J Refrig,2017,38(3):114-118.
[16]张浩.牛血清在细胞培养中的作用与质量要求[J].科技情报开发与经济,2001,11(3):46-47.
[17]S覬ndergaard RH,Follin B,Lund LD,et al.Senescence and quiescence in adipose-derived stromal cells:Effects of human platelet lysate,fetal bovine serum and hypoxia[J].Cytotherapy,2017,(19):95-106.
[18]林常敏,李宇,纪影畅,等.毛乳头细胞培养基血清浓度的探索[J].中国美容整形外科杂志,2008,19(3):218-220.
[19]金亦宏,马欣,周莉薇.国产牛血清的现状[J].中国生物制品学杂志,2004,l7(4):254-256.
[20]吕维敏,黄钢妹,曾叶,等.生物材料冻存中低温保护剂的作用机理及实验研究[J].低温与超导,2014,42(5):12-16.