| The present experiment studied the toxic impact of Aroclor 1254 on mice fetal fibroblast, new bom bovine testis fibroblast, and new born bovine kidney fibroblast from the view of the change of cellular SOD activity , MDA content, and ultra-structure. The effect of feeder cell type, growth factors, and embryo developmental stage on establishment of sheep embryonic stem cell was also studied in present experiment. It was found that:1. Efficient dissociation and culture system were established for mice fetal fibroblast, new born bovine testis fibroblast, and new born bovine kidney fibroblast through associated using mechanic method, trypsinization, and chelated material. There have been no report on in vitro culture of new born bovine testis fibroblast. Based on the culture methods of mice fetal fibroblast, which have been well established and widely used, we adjusted enzyme digest period and media concentration, obtained proper testis fibroblast dissociation method. We also got highly efficient new born bovine kidney fibroblast dissociation technique through enzyme digest step addition on Huang Bin's method.2. Three kinds of fibroblasts appeared different morphology and growth character because of variety of animal and tissue origin. Mice fetal fibroblast attached quickly, most of cells attached 30min after explant, reached quick growth stage in 2d-3d, entered plateau stage in 4d-5d. After confluence, there is no clear margin between cells and only nuclear apophysis could be observed. Cell monolayer was smooth like quiet lake surface. New born bovine testis fibroblast attached slowly, there were enough attached cells only until 24h after explant. They reached quick growth stage in 3d-4d, entered plateau stage in 5d-6d. Confluence cells irregularly arranged, cell monolayer was rugged like lake surface have slight waves. New born bovine kidney fibroblast attached slowest among the three kind cells, majority of cells still suspended in third day after explant, only little attached. They reached quick growth stage in 7d-8d. Confluence cells had typical fibro morphology and arranged irregularly, cell monolayer looks like lake surface have lots of vortex.3. It was indicated that our self-made freeze media was more proper than DMSO media for cryo-preservation the three kind of fibroblast. The media was made for cryopreserve sheep and bovine embryo, include three cryoprotection reagent.4. The three kind of fibroblast had different anti-oxidation capacity. The SOD activity in culture media decreased as following order: mice fetal fibroblast culture media, new born bovine kidney fibroblast culture media, new born bovine testis fibroblast culture media.5. The toxic impaction of Aroclor 1254 on the three kind of fibroblasts was functioned through oxidation damage.6. Firstly Aroclor 1254 promote the production of free oxygen and oxide cell itself; as response, cell then increase synthesis of SOD for releasing free oxygen; when toxic function period increased, SOD activity decreased for the reason of releasing large amount of free oxygen in one hand, in another hand cellular SOD synthesis capacity reduced because of oxidation damage. As the result, oxidation damage was more severely.7. It was found from the ultra-structure change of mice fetal fibroblast, new born bovine testis fibroblast, and new born bovine kidney fibroblast exposed in Aroclor 1254that Aroclor 1254 toxic function exhibited in membrane damage in some extent. When the damage was severe, the typical cellular change was cytoplasm vacuole. The three kind of fibroblasts appeared different sensitivity to PCBs. Mice fetal fibroblasts were very sensitive as revealed the most severe cytoplasm vacuole. New born bovine testis fibroblasts were the second sensitive cell type. New born bovine kidney fibroblasts were the least sensitive cell type among the three kind of cells.8. The study had observed some certain changes in the three different sources of fibroblast exposure to the PCB. Some had quite crassitude nucleolus in their nuc...
|
PDF Full Text Request