Constructing Of Taihu Pig Fibroblast And Preadipocyte And The Differentiation Of Preadipocyte

Taking Taihu Pig as the study materials,we succeeded in establishing a fibroblast cell line by the method of explant culture and a preadipocyte cell line by the method of enzyme digestion.The cell lines were analyzed according to the request for setting up a cell line,like observation on morphology,determination of viability,dynamic growth, analysis of karyotype were carried out.We succeeded in inducing the preadipocyte by 50nmol/L insulin,100nmol/L DEX,0.25 mmol/L IBMX and 100 nmol/L rosiglitazone. Preadipocyte were plated in differentiation medium,the degree of adipogenesis and differentiation were analyzed by their morphological changes and oil red O staining. The main results are as following:1.The fibroblast's morpha of Taihu pig is shuttle shape.There are no morpha diversity between primary fibroblast and subcultured fibroblast.Primary preadipocyte begine to adhere for 3～4h.The preadipocyte' s morpha is typical fibroblast.2.Cell viability of fibroblast and preadipocyte before freezing was 96.5%and 95.4%, after thawing Cell viability was 93.2%and 92.3%.3.Population Doubling Time(PDT) of two kinds of cells were approximately 72h. The growth curve of cultured cell in vitro appeared "S" shape and illustrated that cells growing included latent phase,logarithmic growth phase and stagnate phase.4.The results of chromosome analysis are both 2n=36,XY.5.When preadipocyte differentiated into mature adipocytes,the preadipocytes were changed from shuttle shape to oval or round,in which different size lipid droplets were filled.These preadipocytes replayed their hyperplasia process with higher accumulation of drop lipid.6.The changes of preadipocytes differentiation were observed by Oil Red O staining. The cell lines that constructed in this study accord with the request of construct a cell line.We successfully constructed Taihu pig's fibroblast and preadipocyte cell lines.The way to induce preadipocyte into mature adipocyte was constructed,which is the base for researching differentiation of preadipocyte and give some suggestion to the Taihu pig's breeding.