不同冻存液保存的颗粒性脂肪组织活力及移植成活率分析

The viability and the transplanted survival rate analysis of fat granule in cryopreservation with different frozen conditions

目的研究以基础培养液DMEM高糖和胎牛血清作为冻存基础液是否有助于提高脂肪组织复温后的存活率及移植后的成活率。方法分别用1×高糖DMEM培养基和纯胎牛血清作为配制渗透性玻璃化冷冻保护剂的基础液,将脂肪组织放置-180℃液氮中冻存,于冻存2个月后取出脂肪组织,复温后通过肌酸激酶法、锥虫蓝染色、MTTT细胞增殖实验和异种移植等方法,比较脂肪细胞的存活率、酶活性及增殖能力,并于体内移植4个月后取出移植物,比较移植物的体积、吸收率、脂肪组织结构等情况。结果胎牛血清冻存组脂肪组织复温后的存活率、细胞活性、存活能力及移植后的成活率明显高于对照组(P<0.01)。结论胎牛血清作为冻存基础液能够明显提高冻存脂肪组织复温后的存活率、细胞活性、存活能力及移植后的成活率。

Objective To investigate whether the high glucose Dulbecco＇s Modified Eagle Medium and fetal bovine serum as base fluid are benefit for improve the viability and post-transplant survival rate of frozen adipose tissue. Methods Fat granules was cryopreserved at - 180 ℃ in physiological saline, high glucose Dulbecco＇s Modified Eagle Medium and fetal bovine serum respectively, adding with 8% DMSO. Rewarming at 4 weeks, the viability and post transplantation survival rate of frozen fat were determined by trypan blue staining, ereatine kinase activity assay, MTF assay and xenotransplantation experiments. Results Frozen adipose tissue in fetal bovine serum con- taining 8% DMSO showed an increase in cell viability and improvement in post transplantation survival rate （P 〈0. 01 ）. Conclusions FBS as anessential fluid can significantly improve the cell vitality and post transplanta- tion survival rate of cryopreservation adipose tissue.