共查询到20条相似文献，搜索用时 296 毫秒
1.
ZHANG Yu-ling LIU Feng-jun ZHANG Jing-jing ZHANG Yong 《农业科学与技术》2009,10(6):23-25,32
[Objective]The aim of this study was to explore the technical system of induced expression in vitro of goat mammary gland epithelial cell,and evaluate expression efficiency of mammary gland specific vector and foreign protein at the cell level.[Method]Goat mammary gland epithelial cell transfected by human lactoferrin gene was inducted by culturing in DMEM/F12 medium supplemented with 5 mg/L insulin,5 mg/L prolactin and 1 mg/L hydrocortisone.Supernatant was collected per 6 hours and concentrated.Expression situation of foreign protein were detected by SDS-PAGE and Western blotting.[Result]There was target protein expression in the induced culture medium,which molecular weight was about 42 kD.[Conclusion]The method used in this study can induce goat mammary gland epithelial cell to express foreign gene,it lays a foundation for researching heterologous expression of foreign gene and producing mammary gland bioreactor. 相似文献
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SONG Yu-xuan CAO Bin-yun 《中国农业科学(英文版)》2007,6(1):115-120
To regenerate three-dimensional endometrium in vitro as a novel model for studying the mechanism of implantation of embryos, the luminal epithelial cells and stromal cells of the rabbit uterus were separated and cultured in vitro. The type Ⅰ mouse tail collagen was used as scaffolding material. The stromal cells were inoculated in the type I mouse tail collagen, and the luminal epithelial cells were inoculated on the type i mouse tall collagen to regenerate the endometrium in vitro. The regenerated endometrium was cultured in DMEM-F/12 media containing 100 nmol L^-1 progesterone, 10 nM β-estradiol, and 10% fetal bovine serum （FBS） for 3 d. The media were then replaced with CZB containing 100 nM progesterone, 10 nmol L-1 β-estradiol, and 10% FBS, and the mouse blastulas were co-cultured with it. The results of scanning electronic micrography showed that the epithelial cells on the surface of the reconstructed endometrium were covered with numerous slender microvilli and some epithelial cells protruded pinopodes. After culturing for 12 h with the mouse blastula, the shedding, attachment, and implantation of the blastula were observed. The blastula can escape from zona pellucida and attach to the three-dimensional endometrium and is then implanted into it. This study showed that the reconstructed three-dimensional endometrium can serve as a robust embryo implantation model in vitro. 相似文献
3.
ZHANGGui-xue LIUYan HUPeng-fei 《东北农业大学学报(英文版)》2004,11(2):143-147
The histological observation was experimentally conducted on in vitro cultured mouse embryonic myocardium cells and myocardiumoid cell mass. The mouse embryo tissue were cultured and regular pulsatile myocardiumoid tissue could be found. During in vitro culture, the myofllament bundles in the cell were gradually increasing andstrongly connectted each other with embryonic age and there were loose muscle fibers initially and intercalated discs were close to each other. The lose myofllament bundles were developed in muscle fibers with age and the distance between intercalated discs was enlarged. There were myofilamentoid structure in inactive cells and filament peripherily. 相似文献
4.
LI Meng LI Qingzhan 《东北农业大学学报(英文版)》2007,14(4):345-348
Leptin,a protein hormone produced and secreted predominantly by white adipose tissue,has a critical role in the regulation and coordination of energy metabolism.Leptin is produced in the mammary gland by the fat tissue or by the mammary epithelium.In vitro study has shown that leptin triggers apoptosis in mammary epithelial cells.Mammary gland involution is characterized by extensive apoptosis of the epithelial cells.At the onset of involution,STAT3 is specifically activated.Various studies show that leptin act as a paracrine and autocrin factor to influence mammary epithelial cell proliferation and differentiation.This paper reviewed the function of leptin to the involution of mammary gland. 相似文献
5.
ZHAO Feng LI Qing-zhang 《中国农业科学(英文版)》2007,6(9):1149-1156
This study was to determine the patterns and levels of IGF family members＇ expression during postnatal mammary gland development. The authors investigated the protein expression profile of the major components of the IGF axis in murine mammary glands. All the proteins examined, IGF- Ⅰ, IGF- Ⅱ, and IGF- Ⅰ receptor （IGF-Ⅰ R） were expressed at greatly different levels and displayed unique expression profiles. IGF- Ⅱ and IGF- ⅠR were always expressed at significantly higher levels than IGF- Ⅰ. IGF- Ⅰwas localized in adipocytes as well as the epithelial and stromal compartments, but just distinctly expressed where mammary cells aggregated to form ducts, in virgins. The IGF- Ⅱ was localized only on the basal layer epithelial cell membranes of ducts and alveoli, with a peak level on the initiation of lactation. The higher level of IGF- ⅠR compared with IGF- Ⅰ was also found in adipocytes as well as in the epithelial and stromal compartments, especially during pregnancy and late lactation. The IGF- Ⅰ R pathway was obviously significant for the development of the mammary parenchyma and stroma. Overall, the comparison of the expression profiles of these different proteins would strongly suggest that they were likely to have different functions throughout the mammary gland development, and it also highlighted the potential interactions and coregulation of the members of this axis. It seems that IGF- Ⅱ was the major local modulator rather than IGF- Ⅰ by an IGF- Ⅰ R-independent pathway, especially for initiation of lactation. This study has demonstrated the importance and complexity of the IGF axis during mammary gland development and provides a valuable resource for future research in this area. 相似文献
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《农业科学与技术》2013,(10):1454-1456
[Objective] This study aimed to investigate the histopathological changes in chickens artificially infected with infectious laryngotracheitis（ILT）. [Method] Through ar- tificial injection of allantoic liquid containing ILTV into chickens to induce infectious laryngotracheitis, the clinical symptoms in infected chickens were observed; tissues of trachea, lungs, heart, liver, kidney and spleen were collected from dead chickens, and prepared into paraffin sections, followed by histopathological observation under a microscope. [Results] This disease occured in the inoculated chickens 3d later, with a morbidity rate of 95%. The main symptoms were dyspnea, asthma and coughing up of bloody exudate; yellowish-white pseudomembrane was observed on dissected larynx trachea; swelling, haemorrhage, and further erosion were observed on the in- fected tracheal mucosa, resulting in death of chickens, with the mortality rate of 25%; histopathological observation showed that parenchymal organs exhibited exuda- tive inflammation; swelling, degeneration, necrosis and shedding of epithelial cells were observed; disturbance of blood circulation occured.[Conclusion] This study pro- vides reference bases for the treatment of ILT. 相似文献
7.
ZHANG Yu-ling LIU Feng-jun ZHANG Yong 《农业科学与技术》2009,10(4):27-30,100
[ Objective] The aim was to explore technical system of making single transgenic positive cells become colony cells by amplification culture. [ Method] Fetal fibroblasts and mammary gland epithelial cells of single goat fetus of pBLM-C1 which specifically expressed human lactoferrin were cloned. Single cell colony of single transfection cell was prepared with 3 concentrations of 0%, 50% and 100% conditioned culture media. Transfection cell and non-transfection cell were carried out amplification culture by con-culture, neo gene was as screened gene, genome DNA of transfection cell was detected by PCR method. Chromosome karyotype analysis of single colony cell was tested. [ Result] Compared with non-conditioned culture medium, 100% conditioned culture medium could greatly increase survived rate of single colony cells （ FF： 53.33% vs. 10.00%; MGE： 33.33% vs. 6.67%）. Compared with control, con-culture of transfection cell and non-transfection cell could greatly increase rate of transfection cell single colony after amplification culture （ FF： 53.33% vs. 10.00% ; MGE ： 33.33% vs. 6.67% ）, confluence time of amplification culture was significantly decreased （20 -30 d）. The result of PCR showed that the colony cell obtained by above method contained hLF target gene. The result of karyotype analysis showed that most cloned cell chromosomes were normal. [ Conclusion] The study provides a reliable method for separating transgenic cell, inserting and diagnosing ideal vector, and can save expense and time for transgenic animal production. 相似文献
8.
Jiang Xiao-wen Liu Lin Feng Xin-xin Liu Yan-yan Chen Xue-ying Feng Guo-feng Song Jian-chen Li Yan-hua Yu Wen-hui 《东北农业大学学报(英文版)》2017,24(2)
The aim of this study was to select drugs for prevention and treatment of bovine papilloma in vitro, using bovine papilloma virus(BPV-2) infected mouse mammary epithelial(C127) cell lines as the infectious cell model system. C127 cell shapes changed with BPV infection observed under light microscopy. C127 cell shapes changed with BPV infection observed under light microscopy. PCR was used to confirm BPV infected C127 cells. mR NA levels of matrine to BPV-2 infected C127 cells were detected by RT-PCR. Apoptosis of BPV-infected C127 cells treated with matrine was analyzed by flow cytometry. Results showed that 80 μg· mL~(-1) matrine had a direct virucidal and intervention effect of BPV-2. Matrine inhibited the proliferation of BPV-infected cells. Therefore, matrine might be regarded as a therapeutic drug candidate for BPV induced bovine papilloma disease treatment in future. 相似文献
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GAO Xuejun TONG Huili YIN Deyun ZHANG Li Key Laboratory of Dairy Science Ministry of Education Northeast Agricultural University Harbin China 《东北农业大学学报(英文版)》2008,15(4):15-19
To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the recombinant plasmid pECFP-B by genetic engineering technique.Recombinant plasmid pECFP-B was transfected into dairy goat mammary gland epithelial to detect the bactericidal activity of CecropinB.The expression of CecropinB was also detected.The result of RT-PCR demonstrated CecropinB gene was expressed in transfected cells.CecropinB recombinant plasmid DNA was injected into udders and CecropinB was expressed in mammary gland,exhibiting bactericidal activity to Staphylococcus aureus in vivo experiments. 相似文献
10.
Using embryonic myoblasts to research the formation and de-velopmental mechanisms of skeletal muscle is becoming a research hotspot. This study aimed to establish a method of isolation, culture and identification of my-oblasts in duck embryos. [Method] Pectoral and leg muscle samples were isolated from the embryos of Gaoyou duck at 13 d of hatching, then disassociated with col-lagenase and trypsin and purified via differential adhesion. The isolated cells were cultured in vitro and detected for the expression of Pax7 protein using immunofluo-rescence technique. [Result] Myoblasts were obtained successful y both from pectoral and leg muscles in duck embryos and these cells proliferated strongly and differen-tiated wel . Immunofluorescence staining showed that more than 95% cells could express Pax7 protein. [Conclusion] In summary, we report the successful establish-ment of a complete system for the isolation, purification, identification and culture of myoblasts from duck embryos. 相似文献
11.
[目的]探讨在生化培养箱中进行奶牛乳腺上皮细胞原代培养的可行性。[方法]采用组织块法,在体外进行乳腺上皮细胞的分离培养,探索其在生化培养箱中合适的培养条件。用倒置显微镜观察、细胞爬片吉姆萨染色和角蛋白免疫组织化学染色的方法对培养细胞进行形态学观察和鉴定。[结果]通过倒置显微镜观察发现,上皮细胞呈多角形,长满后呈上皮细胞特有铺路石样。细胞染色可见上皮细胞胞体较大,胞核深蓝色,圆形或椭圆形,核仁清晰可见,一般为2-4个。免疫组化鉴定结果显示,培养的细胞表达上皮细胞特异的角蛋白14和18。[结论]原代奶牛乳腺上皮细胞可以在生化培养箱中成功培养。 相似文献
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【目的】探索一种简单且高效的原代奶牛乳腺上皮细胞的培养方法。【方法】采用组织块培养法、酶消化法培养奶牛乳腺上皮细胞;通过MTT法检测吸光度绘制细胞生长曲线;通过免疫组化法鉴定角蛋白-18的表达。【结果】酶消化法相比组织块培养法获得的细胞具有耗时短,获得量高等优点,MTT法绘制的生长曲线符合一般生物学特性,呈"S"型,奶牛乳腺上皮细胞中角蛋白-18阳性表达。【结论】酶消化法是一种简单且高效的培养原代奶牛乳腺上皮细胞的方法。 相似文献
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体外培养的牛乳腺上皮细胞形态研究 总被引：7，自引：0，他引：7
通过有效的细胞培养方法获得了牛乳腺上皮细胞系,并系统观察了乳腺上皮细胞的长出、贴壁、聚集、迁移、分裂、分化、凋亡等一系列形态变化。结果发现,原代培养的乳腺上皮细胞大多数呈卵圆形,细胞之间连接成片,单层生长,如鹅卵石铺过路面,乳腺上皮细胞和成纤维细胞混生时分区生长,界线明显。传代的乳腺上皮细胞呈岛屿状聚集生长,细胞核呈圆形或椭圆形,核仁2～4枚;在含雌激素的培养液中,细胞出现双核或多核现象,但仍表现一定的接触抑制现象。多次传代后的乳腺上皮细胞含不同的细胞类型,通过光镜观察,部分细胞仍保持较快的分裂增殖能力;部分细胞渐渐分化,出现长形细胞、三角形细胞。上皮细胞增殖分化可形成圆顶型结构,乳腺上皮细胞可产生并分泌乳汁,分泌到细胞外的乳汁流动形成网状结构。 相似文献
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构建pcDNA3.1-hTERT、pcDNA3.1-SV40 T载体,线性化后,共转染荷斯坦奶牛乳腺上皮细胞,研究人端粒酶逆转录酶(hTERT)和猿猴病毒40大T抗原(SV40 T)对荷斯坦奶牛乳腺上皮细胞体外培养的作用,并对细胞进行RT-PCR检测分析及免疫荧光鉴定。结果表明,hTERT和SV40 T在奶牛乳腺上皮细胞中表达可以有效地延长细胞的体外培养时间,增加细胞传代次数,获得的细胞系可以正常表达角蛋白。说明在体外培养的乳腺细胞中共表达hTERT和SV40 T可以有效延长细胞寿命且不影响乳腺细胞特性。 相似文献
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[目的]研究香蕉胚性悬浮细胞系建立过程中非胚性成分的去除方法。[方法]以巴西蕉的花蕾进行诱导培养后,运用孔径在250~500μm的不锈钢筛网进行过滤,并用倒置显微镜对培养物进行实时观测。[结果]运用孔径在250~500μm的不锈钢筛网能够有效去除悬浮细胞系中的褐化坏死组织和分生组织小球,但如果在悬浮细胞系诱导后3 d立即进行去除,则易使之后胚性细胞的增殖变得困难;如果在悬浮细胞系诱导后14 d左右去除,则褐化坏死组织释放的酚类物质很容易损伤并杀死正在增殖的胚性细胞。悬浮细胞系诱发后7 d是一个比较恰当的过滤去除时间点。[结论]在培养的过程中,将悬浮细胞系定期放置在倒置显微镜下进行观察,用移液器能将液泡化的细胞和胚去除。在操作前,需将倒置显微镜放置在超净工作台内,用紫外线长时间照射进行表面灭菌。这样即可以将悬浮细胞系中的非胚性成分过滤去除,又能预防感染。 相似文献
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《东北农业大学学报》2020,(1):57-64
为探讨通草对奶牛乳腺上皮细胞乳糖合成及相关基因表达的影响,采用不同剂量通草提取物处理奶牛乳腺上皮细胞,利用四甲基偶氮唑盐(MTT)法检测乳腺上皮细胞增殖能力,乳糖/半乳糖检测试剂盒(Lactose/D-Galactose (Rapid) Assay Kit)检测乳腺上皮细胞培养液中乳糖含量,实时荧光定量PCR技术检测乳腺上皮细胞乳糖合成相关基因葡萄糖转运蛋白1(GLUT1)、葡萄糖转运蛋白4(GLUT4)、葡萄糖转运蛋白8(GLUT8)、葡萄糖转运蛋白12(GLUT12)、己糖激酶Ⅰ(HKⅠ)、己糖激酶Ⅱ(HKⅡ)、β-1,4-半乳糖基转移酶-1(β-4GALT1)和α-乳清白蛋白(α-LA)基因mRNA表达水平。结果表明,200、400、600μg(生药)·mL-1通草提取物提高奶牛乳腺上皮细胞增殖能力(P<0.05),促进奶牛乳腺上皮细胞合成分泌乳糖(P<0.05),并显著上调细胞GLUT1、GLUT8、HKⅡ、β-4GALT1及α-LA mRNA表达水平(P<0.05),但对GLUT4、GLUT12和HKⅠmRNA表达水平无显著影响(P>0.05)。研究表明,适当浓度通草提取物可提高奶牛乳腺上皮细胞增殖能力,并通过上调乳腺上皮细胞GLUT1、GLUT8、HKⅡ、β-4GALT1和α-LA的mRNA表达,促进乳腺上皮细胞合成乳糖。 相似文献
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采用I型、II型胶原酶和胰蛋白酶混合（1：1：1）消化乳腺组织,并用差速贴壁法纯化培养乳腺上皮细胞,倒置显微镜下观察细胞的形态,并用角蛋白18免疫荧光染色的方法对细胞进行鉴定。结果表明：改良型混合酶消化法能获得较多的细胞团,且12h后细胞团绝大多数已贴在细胞瓶壁,72h后细胞团完全铺开呈单层融合,细胞呈典型的铺路石样。免疫荧光鉴定结果显示,角蛋白18呈阳性反应。因此,应用改良型混合酶消化法能在短时间内获得大量较纯的乳腺上皮细胞。 相似文献
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用过氧化氢(H2O2)建立体外奶牛乳腺上皮细胞的氧化应激损伤模型,研究不同质量浓度茶多酚对氧化应激所致奶牛乳腺上皮细胞损伤的影响。MTT法检测细胞存活率,比色法检测细胞培养液中丙二醛(MDA)含量、超氧化物歧化酶(SOD)和乳酸脱氢酶(LDH)活性,流式细胞术检测细胞凋亡情况,分光光度法检测天冬氨酸特异的半胱氨酸蛋白酶3(Caspase-3)的活性。结果显示:与H2O2损伤模型组相比,20～100μg.mL-1茶多酚处理组的奶牛乳腺上皮细胞存活率、SOD活性显著升高,而MDA含量、LDH与Caspase-3相对活性与细胞凋亡率均下降,其中以100μg.mL-1茶多酚处理组效果最显著,说明一定质量浓度的茶多酚可以缓解氧化应激所致的奶牛乳腺上皮细胞的损伤,提高细胞存活率,抑制乳腺细胞凋亡。结论:茶多酚对乳腺上皮细胞氧化应激损伤具有保护作用,其保护作用可能与降低MDA含量,增强SOD活性及抑制Caspase-3活性有关。 相似文献