Experimental Study Of Preadipocyte Seeded Small Intestinal Submucosa For Adipose Tissue Engineering In Vitro

[Objiective] preadipocytes can differentiate directly into mature adipocytes, and small intestinal submucosa(SIS) is compatible with many sorts of cell.Therefore ,it is feasible to reproduce regenerate adipose through seeding preadipocytes on SIS ,then planting the complex into body,which offer a new approach to treat the problem of adipose defect.The objiective in this study include: to culture preadipocytes in vitro and to study the adhere , proliferation and differentiation of preadipocytes on small intestinal submucosa(SIS) . Methods (1) isolated and cultured preadipocytes from human abdominal adipose tissue in enzyme-digesting method and magnitude the number of cells through subculture. (2) The generation 3 preadipocytes were planted on SIS. Cell adhere determinded by MTT assay under different conditions including weting ways, seeding ways and seeded cell density.(3)The preadipocytes on SIS were divided into two groups .one group were cultured with revulsant and the other without revulsant .the cells in pore plate are treated in the same way .Cell number in different periods were determinded by MTT ,proliferation were showed by the OD—TIME curve .(4) Flow cytometry (FCM) was used to tell cell cell cycle , proliferation index and apoptosis rate of preadipocytes on SIS and pore. (5) Preadipocytes on SIS was seen with the help of histological and Ultrastructural analysis (6) The volume of adipose in preadipocytes induced on SIS was determinded by oil O staining and contrasted withthe volume of adipose in preadipocytes isolated from SIS and those from pore plates. Result weting ways had no effects on the cell adhere but seeding ways did. Draping resulted in more seeding rate (72%) than merging (25%), (p<0.05).The best cell density wasl x 10 5I cm2. (3) The preadipocytes on SIS grew rapidly and the cell number was higher than preadipocytes in pore plates on day 13(p < 0.05). (4) Preadipocytes on SIS differentiated poorly while the preadipocytes isolated from SIS differentiated well as those can on pore plate. (5) Flow cytometry (FCM) showed cell on sis were of higher proliferation index (PI) than that in pore plate. (6) Histology and Ultrastructural showed preadipocytes adjoined tightly SIS and grew well on surface of SIS. Conclusion (1) It is feasible to isolate, culture , proliferate and induced peadipocytes into mature adipocytes in vitro. (2)SIS is very compatible with preadipocytes, so it is expected bio-derived matieral for adipose tissue engineering. (3) However it is necessary to research deeply the positive effect on preadipocyte differentiation and the differentiating potential of the preadipocytes on SIS in vivo.