The Differentiation Of Murine Mesenchymal Stem Cell And Induced Into Adipocytes

Human mesenchymal stem cells are thought to be multipotent cells, which are present in adult marrow, that can replicate as undifferentiated cells and that have the potential to be differentiate to lineages of mesenchymal tissues, including bone, cartilage, fat, tend on, muscle, and marrow stroma. Cells that have the characteristics of human mesenchymal stem cells were isolated from bone marrow of volunteer donors. These cells displayed a stable phenotype and remained as a monolayer in vitro. These adult stem cells could be induced to differentiate exclusively into the adipocytic,chondrocytic,or osteocytic lineages. Individual stem cells were identified that, when expanded to colonies, retained their multilineage potential. multilineage potential have application in fields which are nerval, hematopoietic system, heart diseasea and so on. It is wide used on autografting and allograft, which is easy to be transfected make it become targeted cell to gene therapeutics. it is good methods to difficult diseases. today studies have made models of rabbit, human, murine, baboon, porcin defferation into adipocytes. including AP₂, adipsin, PPAR₂, CEBPn and CEBP/β, CEBP/αlipid-activated transcription factors. meanwhile BMS promote both adipo-progenitors and form ation of adipocytes. Firstly dexamethasone is mainly induce drug, it promote related up-regulation gene expression of adipogeneic and cartilage. the rate is 10%-20% recently years Janderova according to contrast bolting construct project :the medium was changed to Dulbe cco's Modified Eagle's Media (Invitrogen) supplemented with dexa methasone, 3-isobutyl-1-methy xanthine ,insulin . and Scavo indicated that added poli-little concentration insuline growth factor-I can advance adipogeneic. otherwise hypoxia and hypoglycemic agent including to siglitazone appears to exert a potent lipogenic effect independent of PPAR-γ2 maturation pathway. though many projects be reported, the form of percentile have not be determined. if treatment with different dose or conditions, make it differentiating into chondrocytes, osteoblasts and adipocytes, there are quantity is different. some times adipocytes just take possession of 20%-30%, it have no stable condition of differentiation into adipocytes. we observe passage 5 of mesenchymal stem cells which purifed be induced to adipocytes by oil red o staining show orange, and endochylema filled with accumulation of lipids. nulli-coincide 10 fields be observed by Fluor escence Microscopy showed that percentage of adipocytes reach up to 90%, AP(alkaline phosphatase) staining is positive , mac-1, VIII factors antigen staining is negative. that results deplete possible of heterophil granulocyte, macrophage and hematopoietic cells. our expreiments objected to establish the methods of islotion and optimizr t he suitable conditions of induction for mscs differention into adiposities in vitro,according to immunehistochemistry methods to identified rates of induction and Analysis of cytoplasmic lipid, to establish better foundation on histio-engnieering studies of MSCs differentiation into adipocytes on conditions of physiology and pathology.OBJECTIVE To establish the methods of isolation and charact erization of murine bone marrow mesenchymal stem cells, to optimize the suitable conditions of induction for MSCs differentiation into adipocytes in vitro, to explore the possibility of MSCs whether available or not for the adipocyte tissue engineering.METHODS The differential adherence to plastic was employed to isolate MSCs. CFU-F and successive CFU-F cultures were employed to characterize the potent of proliferation and self-renewal of MSCs. The different adipogenic medium was used as induction for the differentiation of MSCs into adipocytes. The differentiated cells were identified by oil red O immunohistochemistry stain.RESULTS The purified MSCs showed the morphology of fibroblasts. It was found that the number of CFU-F formation depended on the planted number of mscs. It showed a good relationship. Small type colony of CFU-F had little potent to re-clone, but almost 90% big type colony of CFU-F had the potent to regenerate CFU-F. The mscs could committed differentiated into adipoGytes induced by different adipogenic medium. But more than 96% MSCs differentiated into mature adipocytes When induced by conbined with dexame thason 1-methy-3 isobutylxanthine, insulin and indomethacin.CONCLUSIONS Our findings showed that we could harvesting the purified MSCs by isolation of differential adherence to plastic, and this MSCs had the potent of proliferation and self-renewal. Moreover, more than 96% MSCs could differentiate into mature adipocytes when induced by combined with dexamethasone, 1-methy-3-isobutylxanthine, insulin and indomethacin. Our findings suggested that MSCs could be valuable for adipocyte tissue engineering.