The Experimental Study Of Transplantation Of Bone Marrow Stromal Cells To Treat Limb Lymphedema

Purpose:The purpose of this study is to identify the curative effect and factors which affect the results of BMSCs transplantation,to explore new methods to treat lymphedema.Methods:1.Operation and radiation were combined to acquire laboratory animals.2.Gain,disassociation,cultivation,marking and assessment of BMSCs by gradient centrifugation and adherent culture.DAPI was added in to mark the third generation cells.3.Transplantation of BMSCs:BMSCs with/or VEGF-C protein were injected into lymphedema limbs.4.Detection to evaluate results of transplantation:①To measure volume change of limbs②To count lymph vessel③Using immunohistochemisty to detect VEGF-C,flt-4 and vWF in lymphedema tissues.④Using RT-PCR to detect VEGF-C,flt-4 and vWF mRNA.⑤Using Western blot to detect VEGF-C,flt-4 and vWF protein.Results:Animal Models are successfully gained by operation and radiation.The achievement ratio was 81%.The swelling and distension of lymphedema.Comparison among different groups,the swelling and distension of lymphedema limbs in four groups was same(P＞0.05) before treatment and 1 week after treatment.But there was great statistical significance(P＜0.01)among all groups 2 weeks after treatment Comparison at different period in the same group:In Group A" there was no statistical significance(P＞0.05) 1 week after treatment and there was great statistical significance(P＜0.01) 2 weeks after treatment.In Group B there was no statistical significance(P＞0.05) 1 week after treatment and there was statistical significance (P＜0.01) 2 weeks after treatment.In Group C there was statistical significance(P＜0.05) after treatment.In Group D there was no statistical significance(P＞0.05) at different period.Micro-lymphatic vessel number:Comparison among the four groups at the same period,there was no statistical significance(P＞0.05) 1 week after treatment between Group B and Group C.But there was great statistical significance(P＜0.01) among other groups.There was no statistical significance(P＞0.05) 2 week after treatment between Group B and Group C.But there was statistical significance(P＜0.01) among other groups.There was statistical significance(P＜0.01) among all groups 3 and 4 weeks after treatment.Using RT-PCR to detect VEGF-C,flt-4 and vWF mRNA:Comparison among different groups,VEGF-C/GAPDH,P＜0.05, FLT-4/GAPDH,P＜0.01,vWF /GAPDH,P＞0.05.Using Western blot to detect VEGF-C,flt-4 and vWF protein:Comparison among the four groups, VEGF-C/β-actin,P＜0.05,FLT-4/β-actin,P＜0.01,vWF/β-actin,P＞0.05.Conclusions:1.Animal Models are successfully gained by operation and radiation.2.BMSCs transplantation can greatly increase new lymphatic vessel,promote reconstruction of the lymphatic return netwoks,to ease and cure the limb lymphedema.3.BMSCs transplantation as one treatment for limb lymphedema is safe:No stem cells marked by DAPI was found 1 week after transplantation.No lymphangioma or hemangioma was found.4.VEGF-C protein injection in local tissue to treat limb lymphedema is effective but poor in long-term effect.BMSCs transplantation combined with VEGF-C protein can do better job.