长大二元杂交猪胎儿成纤维细胞的生物学特性

【目的】研究长大二元杂交猪胎儿成纤维细胞的生物学特性。【方法】采用室温消化法从妊娠30d的长大二元杂母猪胚胎中分离胎儿成纤维细胞,利用MTT法检测第5、10、15、25、35、45、60、70代胎儿成纤维细胞的增殖能力,通过倒置显微镜观察不同代数的细胞形态,流式细胞仪分析细胞周期变化,DAPI染色观察细胞核染色质的形态以及衰老相关β-葡糖苷酶活性的染色测定等方法来判断细胞的衰老程度。【结果】①采用消化液室温消化法成功分离到猪胎儿成纤维细胞,细胞具有典型的成纤维细胞形态。②猪胎儿成纤维细胞于生长初期1～6d呈对数增长,此后进入平台期。【结论】随着细胞代数的增加,细胞的增殖能力虽有所下降,但仍然呈稳定的增长状态,各代次猪胎儿成纤维细胞生长良好。

Study on the Biological Characteristics of Fetal Fibroblasts of the Dual Hybrid Porcine

【Objective】 Study on the biological characteristics of fetal fibroblasts of the dual hybrid porcine.【Method】 A porcine fetal fibroblast cell culture system was successfully established by digestion in room temperature.MTT assay was used to detect the ability of cell proliferation of the fifth,tenth,fifteenth,twenty-fifth,thirty-fifth,forty-fifth,sixtieth and seventieth generation,respectively.Cell morphology was observed by inverted microscope.Cell cycle change was analyzed by flow cytometry.The degree of cell senescence was determined by DAPI staining and senescence-related β-glucosidase enzyme activity stain.【Results】 The results as following: ① Porcine fetal fibroblast cells were successfully isolated using 0.25% Trypsin-0.4% EDTA digestion at room temperature;② Porcine fetal fibroblast cells grew in logarithmic growth at early stage from one to six days,then into the plateau.【Conclusion】 With increases of cell generation,cell proliferation was progressively decreased,but cells were still in a steady growth state.