Antigenic Specificity Of Osteoinductive Allogeneic Rats BMSCs, ADSCs And PMSCs

ObjectiveDiscuss the antigenic specificity of osteoinductive allogeneic rats BMSCs,ADSCs and PMSCs in external environment.MethodsWistar rats of8w SPF inbred line, drawing out three kinds ofmesenchymal stem cells: BMSCs, ADSCs and PMSCs, separating culture theabove cells. The cells growth curve was recorded by MTT method. The3-5generation cells were used to identify cells and orientation for inductingosteoblasts via CD molecular surface markers. The capacity of osteoblastswere detected by the staining of alkaline phosphatase and calcium nodulesdyeing. the influence of the three induced cells to allogeneic WKY rats ofSPF inbred line PUVASP proliferation were detected by applying Transwellco-culture system; positive rate of CD4+, CD8+were measured by the flowcytometric detected; the content of TGF-β1, IL-2were detected by theimmune enzyme reaction monitoring.Results(1) Cell morphological differences: BMSCs cells are elongated, ADSCscells tend to be rounded, PMSCs cells tend to be multi angles.(2) Growthcurve analysis: PMSCs’ latency was shorter, entering a relatively fastlogarithmic growth after passage, entering into the platform4-5d; whileBMSCs and ADSCs proliferation is opposite, logarithmic growth focused between3-5d, Sixth-day later cells into the platform. Three kinds of cells canbe amplificated by the in vitro method for mass culture; Growth curve analysisshows that after being osteoblasts induced the latency of the three cells were1day,after being induced PMSCs entering into the platform3-5days, theBMSCs and ADSCs4-5days.(3) The identifion of CD molecules on thesurface of P5(before osteoblasts induced): the PMSCs CD29+is85%, CD34+is1.33%; BMSCs cell CD29+is90%, CD34+is0.97%; ADSCs cell CD29+is80%, CD34+is0.99%.(4) The cells by alizarin red staining were visible purplecalcium nodules, the result indicate that the three cells can be induced intoosteoblasts; three cells showed a coffee particles after alkaline phosphatasestaining, Kaplow ratings: PMSCs (4points)> BMSCs (3points)> ADSCs(2points).(5) In vitro and allogeneic lymphocyte co-culture test, MTT testshowed three cells are shown to inhibit lymphocyte proliferation, inhibition oflymphocyte proliferation is related to induction time. The analysis showed thatbefore and after three kinds of cells were induced and between three lateralcomparisons of immunogenicity, there was no significance. After cocultruedthe CD4+, CD8+of lymphocytes were decreasedvia the detect of flow cytometry. Co culture supernatant were detected by theImmune enzyme reaction showed that the IL-2expression reduced theTGF-β1expression increased.ConclusionsThe analysis showed that there was no significance in both before andafter three kinds of cells were induced and between the three lateralcomparisons of immunogenicity.