| Background: The application of autologous fat transplantation has been more than one hundred years of history,because of its convenient,non-rejection,safe and effective and other advantages to be accepted,a wide range of applications,has been a lot of scholars of the hot spots.Fat as a high-quality filler,the current clinical application of the biggest difficulty is the high absorption rate after transplantation.Patients need to suffer multiple pains and again high consumption,doctors also increase the medical risk.At present,"a liposuction,multiple injections" is a good way to improve its high absorption rate.In the past,many scholars have reported the article related to fat frozen,through the experiment found that frozen fat still has a high cell activity,to achieve the transplant requirements.There have been many studies on cryoprotectant,and dimethyl sulfoxide has been considered to be an effective cryoprotectant through numerous studies,but its bio-toxicity has been of nearly zero clinical value.In this study,trehalose was used as a cryoprotectant,which was a non-permeable protective agent which could stabilize the biofilm and protein structure.It reduced the damage of the oxidative stress to the organism.It had the effects of antiAnti-cryoprotective effect;there is the role of biological products drying and preservation;in the frozen and resuscitation can make osmotic pressure caused by greatly reduced cell damage.It is important that trehalose is not currently found to be significantly cytotoxic,trehalose has been used clinically for organ preservation solution and eye drops.The conclusion that adipose-derived stem cells are the biggest hotspot in fat transplantation is that the survival rate of transplanted fat has been supported by many scholars.Stem cells are pluripotent stem cells that exist in a variety of tissues in the body with multiple differentiation potentials.Because it is easy to separate amplification,multi-directional differentiation and low immunogenicity and other characteristics,and become the current tissue engineering and regenerative medicine in the seed cells.For the first time,stem cells were transplanted with frozen fat to observe whether the survival rate of postpartum fat transplantation was improved,with the aim of "one liposuction and multiple transplantation".Objective: In this study,the adipose tissue of human granules was mixed with stem cells and transplanted into the dorsal skin of nude mice.The tissue structure,weight and number of blood vessels of the graft were observed to explore whether the combined transplantation of human adipose-derived stem cells and frozen adipocytes Helps the survival of frozen fat cells after transplantation.Methods: The adult female abdominal or thigh fat and separation and purification,we found in the previous study of the better cryoprotectant trehalose,0.35 mmol / L trehalose and other volume added in the adipose tissue,mixed after cryopreservation In liquid nitrogen.After 12 weeks,the third generation of adipose stem cells obtained by extraction culture was mixed with 500 mg: 6 * 105 and transplanted into the left side of group A and group B as the experimental group.Frozen fat was injected into the right side of the back of the nude mice as a control group.The transplantation time of group A was 4 weeks,and the transplantation time in group B was 12 weeks.After 12 weeks,the nude mice were sacrificed.The survival rate of adipose-derived adipose-derived adipose tissue was analyzed by means of graft weight,HE staining,HE staining,oil red O staining,immunohistochemical analysis and number of blood vessels Impact.Results: Cell experiments: 1,adipocyte extraction and culture after 1 day can be observed a small amount of polygonal cells adherent,and then these cells gradually transformed into spindle cells.2,adipose stem cell flow cytometry results: CD29,CD44,CD90 is high expression,and CD34,CD45 low expression.(338.78 ± 30.15)mg,(247.66 ± 27.59)mg,the average weight of the grafts in the experimental group B and the control group were the mean weight of the grafts in the experimental group A and the control group(318.22 ± 17.19)mg,(195.06 ± 10.30)mg.The number of microvessels in the control group was 53.12 ± 2.42 and37.50 ± 1.60,respectively.The results of experimental group B and control group B were 49.75 ± 1.03 and 29.63 ± 1.99 respectively.Conclusion: The survival rate of adipose tissue-treated stem cells after cryopreservation was significantly higher than that of frozen fat transplantation at 4 and 12 weeks. |
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