The Effect Of RMSCs On The Improvement Of Rat Random Flap Survival And Its Possible Mechanism

Objective: To investigate the effect of rMSCs on the improvement of rat random flap survival and its possible mechanism.Methods: Rat bone marrow stromal cells from adult rats were cultured in vitro. To detect the surface antigens ,the labeled cells were analysed with a FACScan flow cytometer. RT-PCR and Western-blotting were used to detect the expression of VEGF in rMSCs. Sixteen SD rats were randomized equally into two groups : rMSCs and control groups,with the former group subcutaneouly injected the rMSCs marked by DAPI during the operation .Ischemic random skin flaps (1.5cm×8cm) were constructed from the rats.Seven days later ,the amount of viable tissue within the flap was measured. After the animals were killed,specimens from the random skin flaps were harvested for immunohistologic evidence of VEGF protein expression and for HE staining to examine the microvascular growth.Results: Rat marrow stromal cells can adhere to the wall and were polymorphic. After P5 culture, 1×1011 cells were obtained. These expanded attached rMSCs were uniformly postive for CD29,CD90,without the expression of CD34,CD45. VEGF can be expressed in the rMSCs cultured in vitro .The marked cells were found in the specimen .The tissue necrosis rate in the rMSCs group was lower than that in the control group(P<0.05),and the average vessel number in the rMSCs group was higher as compared with the control group(P<0.05).Immunohistochemical staining...