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发育至第19期和第28期的鸡胚性腺原始生殖细胞(PGCs)分离提纯后,在DMEM中添加冷冻保护剂DMSO(二甲基亚砜)、EG(乙二醇)、蔗糖进行超低温冷冻保存,比较冷冻保护剂在单独或联合使用条件下对PGCs的冷冻保护效果,复苏后台盼蓝染色测定细胞存活率,体外接种培养、传代。结果表明:(1)第19期PGCs冷冻保存中,10%EG+10%FBS+0.1mol/L。蔗糖条件下细胞存活率最高为(92.20±2.18)%,且与10%DMSO+10%FBS的存活率差异显著(P〈0.05),其余各冷冻保护液间存活率差异不显著;第28期PGCs冷冻保存中,5%DMsO+5%EG+10%FBS+0.1mol/L。蔗糖条件下细胞存活率最高为(92.41±2.82)%,但各冷冻保护液间存活率差异均不显著(P〉0.05)。(2)解冻后体外培养的PGCs,在饲养层和3种细胞因子(I。IF、SCF、bFGF)的共同作用下,可持续增殖,传至第3代的PGCs,PAS染色、AKP染色均呈阳性并保持完整的二倍体核型,仍处于未分化状态。 相似文献
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鸡胚血液中PGCs的分离及原代培养 总被引:3,自引:0,他引:3
在鸡胚孵化48,50,54,56,60h的不同时期,分离血液中原始生殖细胞(PGCs),分离前的浓度分别为0.011%,0.007%,0.006%,0.006%,0.004%,0.001%,分离后的浓度分别为0.245,1.21%,1.02%,0.64%,0.27%,孵化48-54h,血液中PGCs浓度差异不显著,但孵化48h的鸡胚液较少,PGCs绝对量较低。孵化52-54h的鸡胚血液量较大,获取PGCs的绝对值较高,故分离PGCs的时间以孵化52-54h为宜,鸡胚血液中PGCs在未中任何外源生长因子而含10%FCS的TCM-199培养基中体外培养,能够存活3-4d。 相似文献
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曾显成 《江西畜牧兽医杂志》2007,(5):14-15
从表现呼吸道症状、拉黄绿色稀粪为主要特征的发病鹌鹑的脑、脾、肺气管粘液中分离到一株病毒,经HA、HI试验、血清中和试验及动物回归试验,证实所分离毒株为新城疫病毒。 相似文献
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转基因技术在基础理论研究和生产实际中都展示出良好的应用前景。目前,哺乳类与鱼类的转基因技术已相当成熟,并获极大成功,而家禽的转基因技术还十分落后。家禽为体内受精,受精后在卵子周围形成卵白、壳膜及蛋壳,同时受精卵发生快速分裂。受精约20h后产出体外时,胚胎已发育到6000个细胞。因此,对家禽进行早期遗传操作十分困难。尽管如此,国外仍在努力探讨克服家禽胚胎遗传操作的难题,并取得一定的进展。据报道,到目前为止国外实验室已研究的家禽转基因技术主要有:1逆转录病毒感染技术,2DNA显微注射技术3精子介导法[1,2]。其中逆转录病毒感… 相似文献
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一、鹌鹑的孵化 1.种蛋的选择及孵化设备:种蛋要求新鲜,以产出1周左右为宜,种蛋贮藏温度为10℃~20℃,蛋重13~15克。孵化设备可采用立体或平面孵化器。 相似文献
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C. B. L. Mariz J. H. V. Silva J. J. Filho M. R. Lima F. G. P. Costa 《Journal of animal physiology and animal nutrition》2017,101(2):389-400
Four experiments were conducted to estimate the phosphorus and calcium requirements for weight maintenance and weight gain in Japanese quails during their growth phase from 16 to 36 days. Japanese quails aged 16 days were used for estimating the phosphorous and calcium requirements for weight maintenance or weight gain, with these quails composing each reference slaughter group and the others distributed in a completely randomized design, housed in cages of galvanized wire (33 × 33 × 16 cm) that were stored in acclimatized chambers with specific environmental temperatures. The light programme used during the 20‐day experimental period was 24 h of artificial light. Analysis of the data showed that the prediction equations for estimating the phosphorus and calcium requirements for weight maintenance and weight gain of Japanese quails between 16 and 36 days of age were P (g/quail/day) = P0.75*(9.3695 + 7.7397*T) + 9.70*WG, in which P is the phosphorus requirement, and Ca (g/quail/day) = P0.75*(363.99 – 8.0262*T) + 28.15*WG, in which Ca is the calcium requirement, P is BW (kg), T is temperature (°C) and WG (g/quail/day). 相似文献
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对发育至第19期和第28期鸡胚性腺原始生殖细胞(Primordial germ cells,PGCs),用6种玻璃化冷冻液1:10%DMS0+10%EG+10%PVP,Ⅱ:20%EG+10%PVP,Ⅲ:20%DMSO+10%PVP,Ⅳ:10%DMSO+10%EG+0.5mol/L Surcose,Ⅴ:20%EG+0.5mol/LSurcose,Ⅵ:20%DMSO+0.5mol/L Surcose进行冷冻保存。结果:第19期鸡胚PGCs玻璃化冷冻复苏后存活率,在冷冻液Ⅱ与Ⅳ之间差异不显著(P〉0.05),Ⅴ与Ⅵ之间差异显著(P〈0.05),其余各冷冻液之间差异均极显著(P〈0.01)。第28期鸡胚PGCs玻璃化冷冻复苏后存活率,在冷冻液Ⅳ与Ⅴ之间差异显著(P〈0.05),Ⅲ与Ⅳ、Ⅴ之间差异不显著(P〉0.05),Ⅱ与Ⅲ、Ⅳ之间差异不显著(P〉0.05),其余各玻璃化冷冻液之间差异均极显著(P〈0.01)。复苏后接种培养传至第2代的鸡胚PGCs细胞,PAS染色、AKP染色呈阳性并保持完整的二倍体核型。 相似文献
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J. Sales M. Skřivan M. Englmaierová 《Journal of animal physiology and animal nutrition》2014,98(6):1054-1059
Mathematical modelling of the relationships between mineral inputs and outputs would enable the prediction of mineral requirements of poultry under a wide range of conditions. To establish the feasibility of possible modelling of mineral requirements, the current study aimed to describe the individual mineral concentrations of whole bodies of quail over the life cycle from hatching to 70 days of age. Quail were reared indoors without any restrictions that could limit growth. Sampling of birds (n = 6–18) was carried out at 0, 3, 7, 14, 21, 35, 49 and 70 days after hatching. Freeze‐dried samples of whole bodies (digestive contents removed) were analysed for ash, and macrominerals (calcium, magnesium, phosphorus, potassium, sodium) and microminerals (copper, iron, manganese, nickel, selenium, zinc). Ash concentration followed a curvilinear trend, with a maximum of 101.7 g/kg dry matter at 32.77 days. Individual mineral concentrations, expressed as a proportion of ash, were fluctuating over time, with the most prominent changes at 3 days and again at either 14 or 21 days. Dissimilar patterns in individual mineral concentrations resulted that ratios between minerals followed inconsistent patterns over time. Although mineral contents in absolute quantities can be described through modelling over the entire life cycle of the bird, it can be concluded that variable concentrations of individual minerals could complicate further model development. 相似文献
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本研究旨在探讨鸡胚盲肠上皮细胞传代培养条件及其特性,为E.tenella损伤机制及抗球虫药的研究提供体外模型。分别用胰酶-EDTA联合消化和分步消化2种方法分离纯化原代鸡胚盲肠上皮细胞,通过测定贴壁细胞覆盖率选择细胞传代的适宜消化方式;筛选了传代细胞培养液中L-GLN和胰岛素的最佳浓度,通过贴壁差进一步纯化盲肠上皮细胞,探索其合适的培养条件。通过形态学观察、透射电镜、免疫组织化学技术、组织化学技术、糖原染色、流式细胞术的方法对传代细胞特性进行了鉴定。结果表明:胰酶-EDTA联合消化、15min贴壁差纯化除去成纤维细胞,72.5mg.L-1 L-GLN、0.05mg.L-1胰岛素和5%FBS的传代细胞培养液更有利于盲肠上皮细胞的生长;经形态学和透射电镜观察,AKP、Vimentin及PAS染色,所培养的传代细胞具有典型的上皮细胞特征,在传代后24~72h盲肠上皮细胞纯度达95%以上,贴壁细胞覆盖率达85%以上;细胞凋亡测定表明,细胞连传5代,活性较好,第6代细胞凋亡率显著增加,贴壁细胞覆盖率降低。本研究提示用该法传代可获得稳定、数量大、活性高、纯度高的鸡胚盲肠上皮细胞。 相似文献
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为探讨不同细胞因子对水牛原始生殖细胞(PGCs)传代培养的影响,将PGCs分别采用A、B、C、D、E、F和G共7种培养基进行培养,即A组:基础液+10ng/mL白血病抑制因子(LIF)+10ng/mL碱性成纤维细胞生长因子(bFGF);B组:基础液+20ng/mL LIF+20ng/mL bFGF;C组:基础液+20ng/mL LIF+10ng/mL bFGF;D组:基础液+20ng/mL LIF;E组:基础液+20ng/mL LIF+20ng/mL bFGF+20ng/mL干细胞因子(SCF);F组:基础液+20ng/mL LIF+40ng/mL bFGF+40ng/mL SCF;G组(对照组):基础液。将机械法分离的PGCs小集落接种到水牛胎儿成纤维细胞(BEF)饲养层上进行传代培养。结果,原代时,A、B、C、D、E和F组的克隆数目都显著高于对照组(P%0.05),其中E和F组的克隆数目显著高于A组(P〈0.05),而与B、C、D组差异均不显著(P〉0.05);1~8代时,B、C、D、E、F组的克隆数目显著高于A组(P〈0.05);对照组传代数仅为2代,A组为5代,而B、C、D、E和F组均为8代以上。结果表明,在传代过程中,LIF起主要作用,20ng/mL的LIF浓度可以满足水牛PGCs传代培养的需要。 相似文献
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本研究对发育至第19期的鸡胚性腺原始生殖细胞(primordial germ cells,PGCs)分离提纯后,在DMEM中添加冷冻保护剂DMSO(二甲基亚砜)、EG(乙二醇)、蔗糖、PVP(聚乙烯吡咯烷酮)分别组成6种慢速冷冻保护液和6种玻璃化冷冻保护液,进行超低温冷冻保存。复苏后苔盼蓝染色测定细胞存活率,体外接种培养、传代。结果:①慢速冷冻保存中,PGCs在慢速冷冻液V(10%EG+10% FBS+0.1 mol/L蔗糖)条件下复苏后存活率最高(92.20%),且与慢速冷冻液I(10% DMSO+10% FBS)存活率之间差异显著(P<0.05)。②玻璃化冷冻保存中,PGCs在玻璃化冷冻液I(10% DMSO+10% EG+20% FBS+10% PVP)条件下复苏后存活率最高(84.15%),且与其余5种玻璃化冷冻液下复苏后存活率之间差异均极显著(P<0.01)。③培养传至第3代的慢速冷冻复苏后PGCs细胞和培养传至第2代的玻璃化冷冻复苏后PGCs细胞,PAS染色、AKP染色呈阳性并保持完整的二倍体核型。 相似文献