A Comparative Study Of Clinical Application Of PRP Preparation Methods

Purpose: Discussing a method which can safely quickly and highly effective inPRP preparation. Using the rabbit adipose stem cells (adipose derived stem cells,ADSCs) proliferation effect for Tubex and the traditional secondaty centrifugationpreparation of autologous platelet rich plasma (platelet rich plasma, PRP) texting theirbiological effectsMeasure:1The epididymal adipose tissue of the New Zealand whiterabbit was resected． Observing the morphological characristic for the introculture of ADSCs which digestion by the type Ⅰ collagenase.2.Guiding the fourth generation ADSCs to fat and bone.oil red O staining twoweeks later and alizarin red staining three weeks later.3.Cardiac puncture; making platelet-rich plasm with the secondary timescentrifugation;platelet count and calculating platelet recovery.4.Control group (180ul cell+20ulDMEM); The plasma group(180ul cell+20ulplasma) Tubex PRP group(180ul cell+20ul PRP); traditional secondary centrifugationpreparation PRP group(180ul cell+20ul PRP) checking the proliferation in24h48h and72h with Cell-Counting Kit-8.5.Checking the platelet-derived growth factor concentration of the PRP abovewith ELISA.Result:1.We can see adherent cells24hours later and red blood cellwhich can not be adhered. Change the liquid48hours later poaching the impurity withPBS. It is thus clear that so much polygonal cells. Cells spreeding after3days andgradually increasing. Cells formed clusters in5days and it is time to trypsin passaged.Passage cells adhered in4-6hours and breeding a times in3-5days.2. Oil red O staining is positive and and control group is negativealizarin red staining is positive and control group is negative too. 3.The cells number of The platelet count is(260.2±96.8)×109L.and the traditionalsecondary times centrifugation is (1508.8±557.0)×109L;The cells number of Tubexis（3140.0±291.5）×109L12.6times of platelet and2.08times of traditional.4.The result of platelet recovery: calculating the recovery of PRP ofTubex and traditionanl secondary times centrifugation preparation withcalculation formula of platelet recovery: the recovery of Tubex is82.86%and thetraditional is72.54%.5. The tubex and the traditional have the proliferation effect onADSCs in cck-8.There are significant differences among tubex and the control group, the traditional andthe control group. They have the difference of statistics.6. The result of ELISA on the checking of platelet derived growth factor. Thepotency of PDGF-AB of prp in tubex is apparently higher than the traditional and thewhole blood. They have the difference of statistics.Conclusion:1.The cells from the rabbit epididymis digested by a collagenase typewas identified as ADSCs. It have the potential of multi-directional differentiation. Itcan become bone cells and fat cells in the specific environment.2. Using the method of platelet count on the prp which comes from Tubex and Thetraditional two times centrifugation and the potency of PDGF-AB by ellsa, Tubex wascorroborated the more high performance, more quickly and safty method in prp making.3.The PRP of Tubex have higher performance and quicker advantage thantraditional two times centrifugation in prompting the proliferation of ADSCs. Providinga method of cell proliferation in vitro.