| Fibroblasts from embryonic mesoderm is one of the main cells in connectivetissue, which have ability to differentiate, to participate in wound healing, and toinduce osteogenesis and adipogenesis. Under certain conditions, fibroblasts,fibrocytes and mesenchymal stem cells can be transdifferentiated each other. It havebeen reported in many literatures that adipose tissue as endocrine organ are involvedin many physiological and pathological processer, to be widely demonstrated.Moreover, the results in our another study indicated that adipocyte appeared in thecervix during the parturient period and can produce prostaglandin E affecting thecervical ripening and labor. The purpose of the study is to confirm the existence ofadipocyte in the mouse cervix during labor; in vitro culture and identification of in themice cervical fibroblasts and in vitro differentiation of mice cervical fibroblasts and3T3-L1fibroblasts into adipocytes; so as to determine the origin of adipocytes.Cervical samples in mice during labor were collected under sterile conditionsand then cut to frozen sections and stained with Oil Red O; cervical fibroblasts werein vitro cultured and isolated by the tissues explant culture, and identified by means ofthe different sensitivity of a variety of cells to trypsin digestion, cell morphology wasobserved fibroblasts were identified using vimentin antibody immunocytochemistry.3T3-L1fibroblast cels werel cultured, cryopreserved and resuscitated. Isolatedfibroblasts and3T3-L1fibroblast were differentiated in vitro into preadipocytes oradipocytes with containing insulin, DEX, IBMX and ROS, and then identified by OilRed O staining and cellular morphology.The results indicated that preadipocytes or adipocytes also existed in the mousecervix during labor similar to condition in caprine cervix. First generation offibroblasts cultured grew from the explanted tissue piece to the ambient enviroment, and formed a cellular halo in which cellular shape is spindle-like or polygon-like.Celluar staining was positive by fluorescent vimentin antibody detection, so itindicated that fibroblasts have successfully been isolated in vitro. At4th day ofdifferentiation process lipid droplets in a few cells can be seen, and at10th day somelarger lipid droplets in cells began to appear, which showed that some cells have beendifferentiated into mature adipocyte. The survival rate of3T3-L1fibroblasts in controlgroup was76%after recovery. From the differentiation of3T3-Ll fibroblasts toadipose cells, at4th day of the process lipid droplets could be seen in a small numberof cells, and at8th day some larger lipid droplets began to appear in some cells, whichindicated that these cells have been differentiated into mature adipocyte. However, itwas found that the number of differentated cells in culture medium is less in thecontrol group than in the experimental group. The differentiated cells were positiveusing Oil Red O staining,, confirming that lipid droplets appeared in the process ofdifferentiation. In the experiment, the cervical fibroblasts in mice have successfullybeen isolated and differentiated into adipocyes by in vitro culture of primary cells. |
