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71.
类器官(organoids)来源于自组织和自我更新的干细胞,是利用干细胞的自组织特性进行体外3D培养后形成的细胞团,与来源器官密切相关,再现了来源器官的三维细胞结构,并为探索来源器官的发病机制提供了新的模型。类器官系统是由自分泌、旁分泌或邻分泌信号调节下的细胞,或者外源性添加的细胞外基质(extracellular matrix,ECM)底物、小分子和生长因子等衍生而来,这些因素的相互作用创造了一个动态的环境,指导干细胞的自我更新和分化,以及细胞在类器官中的自我组装。诱导多能干细胞(induced pluripotent stem cells,iPSC)重编程方法结合3D类器官工具,使患者来源的类器官作为动物模型和人类临床试验之间的桥梁,是对细胞研究和在体试验的补充。在研究来源器官发育、生物学和病理生理学方面,类器官不仅是一种比传统细胞培养更具生理相关性的体外模型,而且还是再生医学和个性化医学领域中的新模型,有望成为研究营养素、药物、毒物及毒素等的作用机制及药物的筛选、再生医学等领域的重要模型。总之,类器官技术的发展增强了人们对器官和组织生理生化功能的认知。作者对肠、脑、肺脏、肝脏、子宫、卵巢等类器官培养和应用的研究进展进行综述,以期为类器官相关科研及应用提供参考。  相似文献   
72.
以大白菜吉红82、#534DH 群体及#438 群体为试材,研究了利用细胞破碎仪机械提取小孢子与人工挤压提取小孢子对大白菜游离小孢子活力及提取量的影响。结果表明:机械提取替代人工挤压可在短时间内同时提取多份样品,并可快速调整小孢子的培养浓度。收集40 个适期花蕾于5 mL 离心管内,3 000 r·min-1 破碎10 s,将提取的小孢子定容于25 mL NLN-13 培养基,可确保小孢子浓度在适宜培养范围内,并成功诱导胚胎发生。  相似文献   
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ABSTRACT

Aims: To assess the change in body condition score (BCS) during the early and late dry periods and its association with postpartum diseases and milk yield in grazing dairy cows from central Argentina.

Methods: BCS assessments during the dry period, and cow health and milk production records up to 90 days in milk (DIM), were collated for cows from 28 farms at monthly visits between 2007 and 2008. Cows were categorised into four groups; those in Group 1 (n=7,067) maintained or gained BCS during the early and late dry periods; Group 2 (n=2,615) maintained or gained BCS during the early dry period and lost BCS during the late dry period; Group 3 (n=1,989) lost BCS during the early dry period and maintained or gained BCS during the late dry period; and Group 4 (n=5,144) lost BCS during the early and late dry periods.

Results: Cows in Group 1 had reduced odds of having retained fetal membranes (RFM), metritis, and clinical mastitis up to 90 DIM than cows in Group 2 (p<0.001), but the odds of disease were similar to cows in Group 3. The odds of having RFM or clinical mastitis tended to be lower in cows in Group 1 than cows in Group 4 (p=0.08). The odds of cows being culled or dying during the first 90 DIM were lower for cows in Group 1 than for those in Groups 2, 3, and 4 (p≤0.05). Mean accumulated milk yield up to 90 DIM was higher in cows in Group 1 than Group 2 and Group 4 (p<0.001), but was similar to that of cows in Group 3 (p=0.28).

Conclusions and clinical relevance: Cows that lost BCS during the late dry period had increased odds of being diagnosed with several postpartum diseases and had decreased milk yield compared to cows that maintained or gained BCS during the entire dry period. Loss of BCS during any stage of the dry period was also associated with increased incidence of culling or death during the first 90 DIM. These results should raise awareness among dairy cattle producers of the importance of properly managing cow body condition during the dry period, especially during the late dry period.  相似文献   
75.
旨在研究马链球菌兽疫亚种(Streptococcus equi ssp.zooepidemicus,SEZ)烯醇化酶(enolase,Eno)对小鼠肺泡巨噬细胞(RAW264.7)吞噬能力的影响。通过构建原核表达质粒获得重组烯醇化酶(rEno),采用台盼蓝活细胞计数法,判定在不同处理浓度和时间下rEno蛋白对RAW264.7细胞的细胞毒性。将rEno蛋白与RAW264.7细胞共孵育后,用SEZ作用于细胞并检测细胞吞菌数量,判断RAW264.7细胞对SEZ的吞噬活性。进一步通过活细胞稳定同位素标记技术(SILAC)和蛋白质谱分析技术(LC-MS/MS),筛选到RAW264.7细胞中可能与SEZ Eno存在相互作用的候选蛋白。结果发现,10 μg·mL-1 rEno蛋白处理对RAW264.7细胞有明显的细胞毒性,且10 μg·mL-1 rEno蛋白处理RAW264.7细胞2和4 h可显著抑制其对SEZ的吞噬作用(P<0.01、P<0.05)。初步筛选到RAW264.7细胞中动力蛋白激活蛋白亚单位蛋白(dynactin subunit protein 2,Dctn)、整合素α-M蛋白(integrin alpha-M)等17种可能与Eno发生互作的蛋白。本研究获得了rEno重组表达蛋白,发现rEno可减少RAW264.7细胞对SEZ的吞噬,互作蛋白的初步筛选也为进一步揭示Eno在SEZ抗吞噬中的作用机制奠定了基础。  相似文献   
76.
AIM: In order to observe the myocardial differentiation capacity of the dedifferentiated fat (DFAT) cells treated with vitamin C in vitro. METHODS: DFAT cells were dedifferentiated from the mature rat adipocytes with ceiling adherent culture. The DFAT cells of passage 3 were used in the study. Vitamin C and/or neonatal rat heart tissue lysate were added into the culture medium to induce myocardial differentiation for 3 weeks. The cell morphology was observed under microscope. The myocardial-specific markers, such as cTnT, GATA-4 and NKx2.5, were examined by the methods of immunofluorescence, PCR and Western blot. RESULTS: Mature rat adipocytes dedifferentiated into fibroblast-like DFAT cells after ceiling adherent culture. The DFAT cells spontaneously differentiated into cardiomyocyte-like cells under normal culture condition with a low incidence. After treated with neonatal rat heart cell lysate, the DFAT cells became cardiomyocyte-like cells that had bigger size, longer shape and myotubule-structure. The expression of cTnT, GATA-4 and NKx2.5 was remarkably increased at both mRNA and protein levels as compared with the normal cultured DFAT cells. The expression of cTnT, GATA-4 and NKx2.5 was further increased in DFAT cells after treating with vitamin C. No spontaneous beating cell was observed. CONCLUSION: Vitamin C enhances the differentiation of DFAT cells into cardiomyocyte-like cells.  相似文献   
77.
Buffalo mammary epithelial cell,cumulus cell and fibroblasts were transfected by adenovirus vectors and compared their transfection efficiency.293 cells were transfected with pBHGloxdelE13cre and pDC316-eGFP by liposome,the virus was collected and titer was detected.Buffalo mammary epithelial cell,cumulus cell and fibroblasts were exposed to different multiplicity of infection (MOI) of adenovirus vectors.After 72 h,the cells were observed with inverted fluorescence microscope,and transfection efficiency was calculated.When the MOI was 25,50,100,200 and 400,the transfection efficiency of fibroblasts were 0.7%,7.0%,9.0%,12.5% and 34.0%,the transfection efficiency of cumulus cells were 42.5%,55.3%,57.4%,76.0% and 80.0%,the transfection efficiency of mammary epithelial cells were 88.7%,100%,100%,100% and 100%.The results showed that the transfection efficiency of mammary epithelial cell was the best,followed by cumulus cell,and fibroblast was poor.  相似文献   
78.
鸡胚因发育过程清楚,长久以来作为基础和应用科学研究领域重要的实验模型,尤其在鸡胚发育早期绒毛尿囊膜阶段,因其血管丰富,是天然的免疫缺陷宿主,是病理学、药理学和肿瘤学等研究领域的理想实验模型。作者简述了发育早期的鸡胚组织结构,并介绍了鸡胚绒毛尿囊膜在肿瘤研究、血管生成、器官移植、烧伤等疾病机理研究中的应用,以及在鸡胚病理模型基础上进行的抗肿瘤药物筛选的应用。重点介绍了鸡胚及禽类细胞系在病毒繁殖和疫苗生产、治疗性蛋白和单抗生产方面的应用研究进展。多种人源病毒、禽源病毒、支原体等可在鸡胚及禽类细胞上增殖,并用于疫苗生产。作者对常用的禽类纤维原细胞和多能干细胞的发展和特点进行了阐述,并总结了商业化的禽类细胞系来源以及部分易感病毒。鸡胚表达系统能够在目的蛋白特定位点产生人源化糖基,减少目的蛋白对人的过敏反应,且禽蛋廉价易得,可作为生产人用单克隆抗体和治疗性蛋白的合适供体。作者介绍了鸡胚及禽类细胞系在生物医药领域应用的最新进展,并对鸡胚作为动物模型在未来的应用进行了展望。  相似文献   
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80.
CDC25A, TSSK3 and P53 expressionsin vitro in cultured sertoli cells after FSH treatment were studied in order to provide some data for further researches of spermatogenesis. Different concentrations of...  相似文献   
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