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《中国兽医学报》2020,(3)
采用组织块贴壁法分离鸡成骨细胞并进行体外培养,细胞纯化后经姬姆萨、甲苯胺蓝、碱性磷酸酶(ALP)和茜素红染色鉴定;在成骨细胞培养液中分别添加0,10,50,100μmol/L褪黑素,连续14 d测定成骨细胞增殖情况;在第1,3,7,14天测定诱导分化培养的成骨细胞ALP活性,并在第14天检测成骨细胞BSP基因表达水平。甲苯胺蓝染色结果与小鼠胚胎成骨细胞前体细胞系MC3T3-E1一致;ALP染色呈阳性,可见大量黑色颗粒;茜素红染色后矿化结节呈深红色。成骨细胞ALP活性在第3天有所降低,但随培养时间延长,ALP活性逐渐升高;添加褪黑素对细胞增殖的影响差异不显著(P0.05),但在第1天和第14天显著提高成骨细胞ALP活性(P0.05);添加褪黑素提高了BSP基因的相对表达量(P=0.09)。结果表明,利用组织块贴壁法,分离得到鸡颅骨成骨细胞,确定了褪黑素促进成骨细胞分化,且具有时间和浓度依赖性,但不影响增殖。 相似文献
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为了探讨氟对体外培养的绵羊成骨细胞中核因子кB受体活化因子配体(RANKL)mRNA基因表达的影响,进行了绵羊成骨细胞体外培养,分别加入含浓度为0、10-6、10-5、10-4、5×10-4mol/L氟化钠(NaF)的培养液,并于第6天提取细胞总mRNA,用半定量RT-PCR方法分析RANKL mRNA表达水平的改变.结果表明,10-6、10-5、10-4、5×10-4 mol/L NaF均能刺激体外培养成骨细胞RANKL mRNA表达,当氟化钠浓度为10-5及5×10-4 mol/L时,可以明显促进RANKL mRNA表达(与对照组相比P＜0.01).这说明氟呈剂量依赖性地刺激成骨细胞RANKLmRNA的表达. 相似文献
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为了探讨氟化物对反刍动物成骨细胞的细胞周期和凋亡的影响,本研究以初生山羊股骨骨膜为材料分离成骨细胞,接种于含15?S的DMEM培养液,于37℃、5%CO2的培养箱中培养。通过形态观察(Hoechst33342/PI双重染色)、DNA ladder电泳、透射电镜、流式细胞仪等方法检测氟对成骨细胞的调控作用。结果表明,高氟(1.0×10-3及2.5×10-3mol/L)使成骨细胞G0/G1期减少,S期细胞增多,抑制成骨细胞周期由合成DNA的S期向G2M期转化,使细胞停滞在S期。1.0×10-5~2.5×10-3mol/L氟可使DNA断裂,诱导成骨细胞凋亡,其中1.0×10-4mol/L氟所致早期和晚期凋亡率最高,分别为3.33%和2.92%。本试验证实氟可诱导成骨细胞凋亡。 相似文献
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《畜牧兽医学报》2018,(12)
成骨细胞在骨代谢过程中发挥成骨作用,笔者制备羊骨髓肽钙螯合物(MP-Ca),探究其对体外培养成骨细胞的增殖及活性的影响,并将其效果同羊骨髓肽(MP)进行比较。制备的MP-Ca经傅里叶红外光谱仪进行扫描鉴定;从新生SD大鼠颅骨中分离纯化的成骨细胞经碱性磷酸酶(ALP)和茜素红染色鉴定;在培养液中添加不同浓度的受试物,CCK-8法确定最佳作用浓度。以17-β雌二醇(10~2μg·mL~(-1))为阳性对照,通过测定各组细胞OD_(450 nm)和ALP、骨钙素(BGP)的含量,考察MP、MP-Ca在最适作用浓度(10~3μg·mL~(-1))下对成骨细胞增殖和活性的影响。结果表明,MP和MP-Ca的红外光谱图未完全重叠,吸收峰发生了明显的位移变化,说明MP与CaCl_2反应形成了MP-Ca。分离的成骨细胞经ALP染色呈阳性,胞内可见大量黑褐色颗粒;茜素红染色后细胞呈深红色,并形成钙化结节。MP-Ca处理组的OD_(450 nm)和ALP、BGP水平均显著(P 0. 05或P 0. 01)高于MP处理组,但均显著(P 0.05或P 0. 01)低于雌激素组。综上,尽管效果不及雌激素,MP-Ca和MP均可促进体外培养成骨细胞的增殖并增强其成骨活性,效果MP-Ca优于MP。 相似文献
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本文旨在探讨不同浓度亚硒酸钠溶液对体外培养的小鼠成骨细胞的形态、增殖及活力的影响。以出生24h的昆明小鼠颅骨为材料,通过酶消化法来分离成骨细胞,并分别接种于不同浓度的亚硒酸钠培养液(0.00、0.05、0.1、0.2、0.4、0.6、0.75、1.0 mg/L)中,倒置显微镜下观察成骨细胞的形态变化,采用MTT法测定细胞增殖率,结晶紫染色法测定细胞活力。48 h内,浓度高于0.4 mg/L时亚硒酸钠抑制细胞增殖且降低细胞活力,而低于0.2mg/L时亚硒酸钠可促进细胞增殖,增强了细胞活力。结果显示硒对成骨细胞的增殖和活力呈双向调节,硒作用呈浓度剂量依赖性。 相似文献
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采集奶牛卵巢表面卵泡中的卵丘-卵母细胞复合物,通过胰蛋白酶消化获得卵丘细胞,分别利用对照组DMEM/F12培养液和4个试验组McCoy's5a培养液对卵丘细胞进行原代及传代培养.试验1组用McCoy's5a基础培养液,试验2组在McCoy's5a基础培养液中添加29 ng·mL-1的睾酮,试验3组同时添加29 ng·mL-1的睾酮和15μmol·L-1的绿茶多酚,试验4组在第3组基础上添加转铁蛋白(5 μg·mL-1)、硒(5 ng·mL-1)和胰岛素(10μg·mL-1),构建成优化培养液(Optimized McCoy's5a).结果表明,试验4组优化培养液所培养的卵丘细胞的形态和增殖规律符合正常细胞的一般规律,染色体核型正常,其细胞内SOD、GSH含量及冷冻解冻后的存活率均显著高于对照组,优化培养液可以代替含血清的DMEM/F12培养液用于奶牛卵丘细胞的体外培养. 相似文献
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《中国兽医学报》2020,(2)
脂联素受体激动剂(AdipoRon)是人工合成的与脂联素(ADPN)有相似作用的口服活性小分子,研究显示ADPN可调节成骨细胞生长和分化,但AdipoRon是否具有类似的功能目前鲜见报道。本试验主要研究AdipoRon对鸡成骨细胞的影响。从14日龄鸡胚额骨中分离获得成骨细胞,细胞培养第4日,分别添加100,200 mg/L AdipoRon处理成骨细胞,处理72 h后,MTT法检测细胞增殖活性,并进行碱性磷酸酶(ALP)染色。Real-time PCR检测脂联素受体1(AdipoR1)、脂联素受体2(AdipoR2)、成骨细胞成熟标志基因骨钙素(osteocalcin,OC)、Ⅰ型胶原α2链(alpa2 of type I collegen,COL1A2)、细胞凋亡相关基因Caspase-3、Bcl-2及Bax的基因表达量,计算Bcl-2与Bax基因表达量的比值。结果显示,100,200 mg/L AdipoRon处理后的成骨细胞正常形态消失,细胞数量减少,体积变小,细胞核明显固缩,细胞存活率极显著降低(P<0.01)。AdipoRon可增加成骨细胞中AdipoR1、AdipoR2、OC、Bcl-2、Bax和Caspase-3的表达量(P<0.05),并呈剂量依赖性,但对COL1A2和Bcl-2/Bax的表达无显著性影响。结果表明,100,200 mg/L AdipoRon均可促进鸡成骨细胞AdipoR1/2的表达,且能抑制成骨细胞的增殖,促进成骨细胞的成熟及凋亡。 相似文献
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本研究旨在观察不同代次骨髓间充质干细胞(BMSCs)和脂肪间充质干细胞(ADSCs)体外培养的生长特点和体外诱导成骨能力。通过密度梯度离心和贴壁培养法分离培养大鼠骨髓间充质干细胞和脂肪间充质干细胞,用含地塞米松、抗坏血酸、β-甘油磷酸钠的培养液定向诱导传代细胞向成骨细胞分化,并利用茜素红染色、碱性磷酸酶染色及PCR方法检测成骨细胞。结果表明骨髓及脂肪间充质干细胞呈成纤维细胞样生长,增殖能力强,生长迅速。第5、10、15、20代BMSCs及ADSCs经诱导培养后茜素红染色呈阳性并且出现"矿化"、碱性磷酸酶活性强,随着细胞代次的递增,诱导后细胞碱性磷酸酶活性呈递减趋势;诱导后的两类细胞传代后细胞仍能继续分化,并形成正常的"矿化"结节,且碱性磷酸酶染色均弱于初次诱导。结果提示,BMSCs及ADSCs易于分离培养及体外扩增,诱导条件下成骨能力强且成骨细胞传代培养仍具有成骨能力,适合作为再生医学骨组织工程的种子细胞。 相似文献
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试验旨在研究不同卵母细胞收集方法及添加半胱氨酸、肝素钠对黄牛卵母细胞体外成熟及体外受精的影响。以黄牛为研究对象,采用两种方法(抽卵法和割卵法)抽取卵泡中的卵母细胞,比较两种方法获取的卵母细胞成熟率,结果发现,抽卵法获得的卵母细胞成熟率显著高于割卵法(P0.05)。将获取的卵母细胞分为4组:A组(对照组,只添加基础成熟培养液)、B组(基础成熟培养液+200μmol/L半胱氨酸)、C组(基础成熟培养液+20μg/mL肝素钠)、D组(基础成熟培养液+200μmol/L半胱氨酸+20μg/mL肝素钠),结果发现,D组的卵母细胞成熟率显著高于A、B、C组(P0.05),B、C组间卵母细胞成熟率无显著差异(P0.05),但两组卵母细胞成熟率均显著高于A组(P0.05);A组卵母细胞卵裂率均显著低于B、C、D组(P0.05),B、C、D组间卵母细胞卵裂率无显著差异(P0.05);D组囊胚率显著高于其他3组(P0.05)。结果表明,抽卵法获得卵母细胞效率显著高于割卵法,肝素钠及半胱氨酸对黄牛卵母细胞体外成熟和体外受精都有促进作用,且同时添加两种物质对体外成熟的效果更佳。 相似文献
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魏静 《四川畜牧兽医学院学报》2009,(4):28-32
在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。 相似文献
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本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。 相似文献
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以国际标准强毒Ｒ株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离ＭＧ,并收集感染鸡所产蛋分离ＭＧ。结果表明,人工感染４８小时后上、下呼吸道及肺已被全面感染,９６小时气囊已被感染,１２０小时输卵管已能分离到ＭＧ,卵巢始终分离不到ＭＧ。人工感染鸡自１４４小时便能在其所产蛋中分离出ＭＧ。药物治疗能在７２小时内消除感染,油乳剂苗则需２４天后逐渐降低蛋内ＭＧ分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内ＭＧ,但以研制的种蛋浸泡剂药浴效果为最好。 相似文献
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REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air. 相似文献
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Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF
Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo. 相似文献