研究利用玻璃针显微分离了巴马小型猪外周血淋巴细胞有丝分裂中期的1号染色体，然后将显微分离的染色体进行简并寡核苷酸引物PCR（DOP—PCR）扩增，通过PCR技术和荧光原位杂交（Florescence in situ hybridization，FISH）技术对扩增产物来源进行鉴定后，将DOP—PCR产物与pMD18-T载体连接、转化以构建1号染色体微克隆文库。结果表明：DOP—PCR扩增产物与猪的1号染色体DNA同源，并且得到了插入片段为100～600bp的巴马小型猪的1号染色体微克隆DNA文库。
[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying biological mechanism of embryo implantation and uterine diseases. [Method] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique were used to isolate BESC and BEGEC, then immunocytochemical method and TRYPAN-Blue assay were used to determine the purity and survival rate of isolated cells. [Result] The BESC and BEGEC were successfully isolated and cultured while immunocytochemical method and cell count method demonstrated that the purity was over 90%. The result of TRYPAN-Blue assay shown that survival rate of BESC and BEGEC was 91% and 78% respectively. [Conclusion] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique could isolate BESC and BEGEC with high purity.
