| Background Adipose-derived stem cells(ADSCs)were extracted from adipose tissue by Zuk.It has been discovered that adipose-derived stem cells are able to get easily,adipose tissue can be induced to differentiate toward adipose cells, cartilage cells,muscles cells,which illustrates the potential of differentiation and cross-lineage differentiation.After a series of experiments,we find that adipose-derived stem cells could promote the cell growth and be ability to repair themselves.Around the adipose tissue,ADSCs could Vitro from stem cells induced to differentiate into adipose tissue, It has the strong self-renewal capacity.Adipose-derived stem cells are able to Improve some ploblems,sush as facial defects and breast.Cosmetic doctors concerned that their development are widely used in Clinical Practice.However,after the adipose tissue graft,due to Infection,clinicians realize that adipose tissue could Survive a little as well as Patients will be dissatisfied.these reasons restrict its development,the low rate of survive lead to the demand of certain trouble.Platelet-rich plasma(PRP) could require from fresh blood.There are many methods to require PRP at home and abroad,Most scholars have chosen centrifugation to get PRP.In a certain period of time,PRP could enhance the rate of survive,yet,we have no the specific mechanisms.Anx A1 gene and PPARγ gene could promote stem cells Proliferation and enhancethe autologous fat transplantation,In a certain period of time,we examine the proliferation level of Anx A1 gene and PPARγ gene by adding PRP into ADSCs,.Objective:The purpose of the experiment is to discover that PRP has effects on promoting Anx A1 gene and PPARγ gene in rabbit ADSCs by adding PRP into rabbit ADSCs.Methods:1. ADSCs were taken from epididymis adipose tissue in a New Zealand rabbit,stripped out the envelope, removed the fat pad and then identify them by Morphology and inducing differentiation, placed in incubator,cultivated them to the fourth generation by Trypsin Digestion.2. Extract 20 ml of rabbit arterial blood in a New Zealand rabbit,prepare platelet-rich plasma(PRP) and platelet-poor plasma(PPP) through traditional secondary centrifugation.3. Experiments were divided into three groups,put purified ADSCs into PRP(experiment), into PPP(comparison) and into a blank groupe with 5% nutrient fluid separately cultured for 24 hours, 48 hours and 72 hours in incubator.4. To obtain overall RNA in every group, Detection the target of PPARγgene and Anx A1 gene Changes in gene expression level by RT-PCR.Results:1. Observed by a microscope,the original ADSCs were ovally shaped,after 4-6 hours the cells started to stick to the edge and to become long shuttle shape in 25cm2 culture bottle.48 hours later, the cells become Short stick and long shuttle shape.Replacing liquid for several times,by culture and subculture to the third or fourth,the purified cells are consistent with the morphological characteristics of stem cells.2. Cloloring them by oil red O and alizarin red namely successfully demonstrates theyare adipose cells and osteocyte,as well as their ability of multilineage differentiation.3. compared with the other two groups,Anx A1 gene and PPARγ gene in PRP group,which expression was significantly increased by RT-PCR.Conclusion:It can be proved that the cells are rabbits adipose-derived stem cells(ADSCs)by morphologic analyzing and multilineage differentiating the cracked and in vitro cultivated cells.Prepare platelet-rich plasma(PRP) can improve the multiplication of ADSCs and enhance the promotion of Anx A1 gene and PPARγ gene... |
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