Effects Of Intravenous Umbilical Mesenchymal Stem Cells Injection On Growth,Development And Serous Biochemical Traits In Kazakhstan Lamb

The process of isloation and cultivation of the sheep’s umbilical mesenchymal stem cells was established through observed biological characteristics of isolated, purificated and cultivated sheep umbilical mesenchymal stem cells (MSCs) in vitro by the method of Mix-enzyme co-digestion and Pancreatic enzyme digestion.Cells were gained from fresh sheep umbilical organization without arteriovenous which was sheared and added mix-enzyme to digesting for24h. From observing the cells morphology after screening to wipe impurity off by Trypsin digestion, cell growth curve of the first, the fifth and the tenth generations was drawed. Osteogenic differentiation capacity induced was detected by histochemical staining(Alizarin Bordeaux staining) in vitro and adipogenic differentiation capacity induced is detected by Oil Red O staining method in vitro. The result shows that there is not obvious change in morphology and proliferation capacity after10generations continuous cultured. Osteogenic cells induced in vitro for21d was proved that there has been induced osteogenic differentiation capacity by von Kossa stained calcification nodules of skeleton cells in cell culture plate, in the meanwhile, adipogenic cells induced in vitro for14d was proved induced adipogenic differentiation capacity by fat droplet in cell culture plate. It is confirmed by experiment that there are differentiation capacity of sheep umbilical mesenchymal stem cells by cultured and expanded in vitro which is similar to BMSCs.Second experiment:Kazakhstan sixty healthy sheep were selected with conditions of be born for about one month and similar weight, then they were divided into two groups(each group thirty sheep, included fifteen male sheep and fifteen female sheep). The difference between two groups are not significant in weight and body measurement. Sheep of experimental group based on randomized approach were injected in jugular vein with isolated and cultured heterogenous umbilical mesenchymal stem cells (5×105stem cells/per kg) from healthy Kazakhstan female sheep umbilical cord.Sheep of control group were injected equivalent physiological saline solution. It was the same of sheep grain ration recipe, nutrition and management between experimental group and control group in testing duration from January to July. Everytime before injected, sheep without eating were tested some indexes such as weight, height, length, chest circumference, circumference of cannon bone and hip width. Serum was collected gather to assay IGF-1, TGF-β1, bFGF, BMP-7, TGF-a, GH, Glu, TP, Alb, TC, TG, LDH, ALP, AST, T-AOC, SOD and MDA levels.Results of second experiment:average weight of experimental group rather significant exceeded control group (P<0.05) in three months, four months and five months meanwhile difference was very significant (P<0.01) in sis months and seven months.In whole duration of test, there is rising trend of both experimental group and control group in height, length, chest circumference, circumference of cannon bone and hip width.These indexes of experimental group were mild higher than control group from the second month, but the difference was not significant (P>0.05). It was tested that growing and developing rate of Kazakhstan sheep were very significant increased and production performance was effective increased and stress effect was reduced by injecting5×105umbilical original mesenchymal stem cells per months per kilogram.Results of second experiment:average weight of experimental group rather significant exceeded control group(P<0.05) in three months, four months and five months. Difference of exceeded degree was very significant P<0.01)in six months and seven months.In whole duration of test, there is rising trend of both experimental group and control group in height, length, chest circumference, circumference of cannon bone and hip width.These indexes of experimental group were mild higher than control group from the second month, but the difference was not significant (P>0.05). It was tested that growing and developing rate of Kazakhstan sheep were very significant increased and production performance was effective increased and stress effect was reduced by injecting5×105umbilical original mesenchymal stem cells per months per kilogram.There was a strong anastomosis between serous IGF-1level and curve of average month growth rate of both experimenttal group and control group in corresponding period with positive correlation in the two groups after injected umbilical mesenchymal stem cells. Serous IGF-1level of experimental group was showed significant higher than control group (P<0.05).Serous GH level and IGF-1level had negative correlation in experimental group compare to control group in corresponding period, but difference of serous GH levels in two groups was not significant (P>0.05).In whole testing period, serous bFGF levels in experimental group were all higher than control group, especially in difference was significantin two months, four months and seven months (P<0.05). There is positive correlation between serous BFGF of experimental group and average increasing weight per month. It is showed declining trend of serous bFGF in every growing and developing fastigium, yet it is rising trend in growing and developing lag phase. There is declining trend of serous BMP-7level in experimental group and control group in fastigium of average increasing weight per month, and significant difference of serous BMP-7level in experimental group was lower than control group (P<0.05). The difference of serous GH level between the two groups was not significant (P>0.05) and so was within each group in different months (P>0.05).It is tested that the serous IGF-1、TGF-β1、bFGF、 BMP-7、TGF-α and GH levels in sheep were direct or indirect effected by nest regulation of stem cells after injected umbilical mesenchymal stem cells.In whole duration of test, serous Glu level in experimental group which had been injected umbilical mesenchymal stem cells was higher than control group, but difference is not significant (P>0.05).Amount of serous TP had increased significantly (P<0.05) in experimental group, and the increasing level was very significant (P<0.01) higher than control group in three months and seven months. Serous Alb、TC and TG level of experimental group were higher than control group, but difference was not significant (P>0.05). Serous ALP level of experimental group is higher than control group in the whole testing time, difference was very significant (P>0.05) in four months and six months. Serous ALT and T-AOC levels in experimental group were all higher than control group, but difference was not significant between two groups, either was difference within each group in every months term (P>0.05).The higher level of serous SOD in experimental group indicated that serous SOD level had been brought rising trend by injecting stem cells. Trend of quantity was significant in two months and four months (P<0.05).There is a lower quantity of serous MDA in experimental group, but the difference was not significant between two groups (P>0.05)Experiment results indicated:synthesize and metabolic energy of experimental sheep has been increased by growth factor which is from secretion itself and paracrine of stem cells nest regulation after transplanting umbilical mesenchymal stem cells.So was growth, development and immunity of tested sheep.