【摘要】 用组织块培养法获得山羊乳腺细胞原代培养物,并根据山羊乳腺成纤维细胞与上皮细胞对胰蛋白酶的敏感性不同将二者分离纯化。对细胞形态进行光镜观察发现:纯化的山羊乳腺上皮细胞传代至第15代时其生长仍正常。纯化的乳腺上皮细胞增殖可形成圆顶型结构,呈乳头状,称之为乳球体;乳腺上皮细胞可产生并分泌乳汁。山羊乳腺上皮细胞含不同的细胞类型,大多数上皮细胞呈短梭形或多角形,呈蜂窝状;部分细胞呈圆饼状,体积较大;部分细胞呈长形。采用电穿孔法将绿色荧光蛋白(EGFP)基因转化体外培养的山羊乳腺上皮细胞,荧光显微镜下观察到EGFP基因的表达。更多还原

【Abstract】 Primary culture of goat mammary gland cells was achieved by outgrowth of migrating cells from fragment of tissue. The fibroblast and epithelial cells were purified according to their different sensibility to trypsin. And the characteristics of the goat mammary epithelial cells were observed by light microscope. The results showed as follows: The purified goat mammary epithelial cells kept normal characteristics until 15th passages. The purified mammary epithelial cells propagated and formed dome-like construction which looked like nipple and was called milk orb; The mammary epithelial cells could produce and secret milk. The goat mammary epithelial cells contained different cell type, majority of the cells were short shuttle-like or guboidal, they looked like beehive; part of the cells looked like big round flat cakes; part of the cells were elongated. The EGFP gene was successfully transfected into the goat mammary epithelial cells using electrotransfection and the expressing of EGFP gene under fluoroscope was observed.更多还原