【作者】 丰艳妮；

【导师】 靳亚平；

【作者基本信息】 西北农林科技大学 ， 临床兽医学， 2006， 硕士

【摘要】 本试验建立一个子宫内膜腺上皮细胞的分离培养体系,探讨了性成熟山羊和羔羊子宫内膜腺上皮细胞的分离纯化方法,并进行其与PBMC的共培养,通过MTT法检测培养的细胞对PBMC活性的影响。为阐明动物不同生殖生理时期内分泌和免疫系统的相互影响提供试验依据,获得了以下结果:1.进行山羊子宫内膜消化试验以及腺上皮细胞分离纯化试验,证明了当用2 g/L的胶原酶于37℃消化4 h时,可以获得大量的细胞,利于细胞的分离。将消化的组织液液过200目滤网后,利用低速离心和反复沉降的方法,得到内膜腺上皮细胞经免疫组织化学鉴定,其纯度达95%以上。2.比较性成熟山羊和羔羊的子宫内膜腺上皮的生长特性,证明:不同时期的腺细胞其生长情况有所差异,羔羊的生长速度较快。3.建立了一种新的共培养方法,并将其与猪PBMC进行共培养。显示,这种方法可满足腺上皮细胞与PBMC相互作用研究的需要;与其他几种共培养方法比较,操作简单。4.证明子宫内膜腺上皮细胞和猪PBMC共培养以及条件培养液均能显著刺激PBMC的增殖;腺上皮细胞能够抑制PHA-P对PBMC的增殖作用。提示子宫内膜腺上皮细胞可影响PBMC的活性,参与了子宫免疫平衡的调节。本试验对子宫内膜腺上皮细胞的分离培养条件的探讨,所得的腺上皮细胞纯度达95%以上;同时,将其与猪PBMC共培养,发现子宫内膜腺上皮可直接影响PBMC。更多还原

【Abstract】 The experiment established the isolated culture system of endometrial epithelioglands, approached the method of separating and purifying the endometrial glands in sex-maturity goats and lambs, moreover, co-culture with PBMC. And we detected the influence of glands to PBMC by MTT. Provide the experimental proof to the interaction between endocrine in different physiological period and immune system,the results as follow:上1. Carred out the experiment of digesting the endometrium and separating the gland cells, proved that when digesting 4 h by collagenaseⅠat 37℃, can get enough cells and be profit for the separation. The digestion was passed over the 200 sieve, purified the cells after gentle centrifugalization and natural sedimentation. Identified the cells we had by immunohistochemistry, the purity rate endometrial glands of is more than 95%.2. Compared the growth characters of the endometrial epithelioglands in sex-maturity and lambs, proved that in different period the growth of gland cells were distinguished and the gland cells in lamb were grew more faster.3. Established a new co-culture method, and co-culture the epithelioglands with PBMC of swine. The result showed that the method can satisfy the requirement of interaction between the two cells. Compared with other method, it is easy to operate.4. Proved that the epithelioglands and conditioned medium both can proliferate the PBMC of swine, and the gland cells could inhibited the proliferation of PBMC which was induced by PHA-P. It suggested that the endometrial epithelioglands could influence the activity of PBMC, and participate the regulation of uterus immune balance.The experiment approached the condition of isolated culture the endometrial glands in goats, the purity rate of cells we had gotten was more than 95%, and co-culture with PBMC of swine, indicated that the gland cells could affect the PBMC directly.更多还原