Isolation, Identification, Induction Of Porcine Adipose Mesenchymal Mtem Cells And Research In Breast Reconstrution Model Combined With Expander

Objects:1. To segregate adipose mesenchymal stem cells (AMSCs) from subcutaneous f atty tissue of wu-zhi-shan minipig. The cells were passaged by traditiona1methods, frozen storage and thawed by generalprocedure after cultured i n vitro. Then to check the surface maker and differentiation of the cell s;To put the tissue expander in abdominal subcutaneous of wu-zhi-shan mini pig, regulate the shape of model, set up a ideal breast shape;To invetigate the differentiation of locall cells,explore the potention of adipose mesenchymal stem cells (AMSCs) in breast reconsruction.Methods:1. Isolation of the primary adipose mesenchymal stem cells (AMSCs):the pr imary adipose mesenchymal stem cells (AMSCs) were isolated from wu-zhi-sha n minipig esenchymal fatty tissue by collagenase I digestion method, cult ured in primary medium. Subcultured and frozen in traditonal way to inspe t the stability. Check the antigen maker by flow cytometry analysis. Detec te the differentiation into adipose of the cells which were cultured in the adipose-inducing medium, exam the differentiation into bone of the c ells which were cultured in the bone-inducing medium;2. To set up the animal model:put the desgined tissue expander under the a bdomianl skin after anaesthesia of zu-zhi-shan minipig. Adjust the shape b y disjunctive injection though the catheter;3.To detecte the results of differentiation in local part:pump ot the no rmal salin (NS) from the expander though the catheter, inject the adipose mesenchymal stem cells (AMSCs).observe the change of shape by eyes, check the differentiation of cells by immunohistochemisty technology;Results:1.we can succesfully isolated the primary adipose mesenchymal stem cells (AMSCs) from fatty tissue by collagenase I digestion method. The stem cel ls were negative for CD34and CD45, postive for CD44and CD105. Meanwhile, t he cells were positive for oil red0staining and alizarin red staining;2. We can set up a breast reconstruction succesfully by injection normal s alin (NS)discontinuously, adjust the shape of the expander by the injectio n volum and time;3. the local cells were much more in experimental group than control group by immunohistochemisty technology.While the changes observed by eyes we re not visibly.the cells can be used in breast construnction.Conclusion:1. The porcine primary adipose mesenchymal stem cells (AMSCs) were isolat ed from wu-zhi-shan mini-pigfatty tissue,could becultured in vitro.the ce11s were also induced to adopogenic and octeogenic differentiation in vir o in special medium. The cells posssess the ability of self renewal and m ulti-directional differentiation, especially conveyde the maker.2. The experimentation on animals revelated the ability of multi-directio nal differentiation to the microenvironment which produced by the adipos e mesenchymal stem cells (AMSCs);3. Porcin adipose mesenchymal stem cells (AMSCs) will be the new idol cell s for breast reconstruction, but there was a long way to explain the spec ific mechanism and safety.