籽鹅不同等级卵泡生殖相关激素受体基因的表达

目的：检测籽鹅不同等级卵泡不同生殖相关激素受体mRNA的表达。方法：选用1只8月龄产蛋期籽鹅,处死后分离各级别卵泡（F1、F2、F3、F4、F5、SYF、LWF）,采用实时荧光定量PCR方法检测促卵泡激素受体（FSHR）、促黄体生成素受体（LHR）、雌激素受体α（ER α）、雌激素受体β（ER β）、生长激素受体（GHR）和胰岛素样生长因子结合蛋白-1（IGFBP-1）6种生殖相关激素受体mRNA的表达量变化。结果：SYF和LWF中LHR、GHR和IFGBP-1 mRNA表达量较低,而F1-F5级卵泡中表达量较高;SYF和LWF中ER βmRNA表达量较高,而等级卵泡中表达量较低。结论：本实验研究了不同等级卵泡FSHR、LHR、ER α、ER β、GHR和IGFBP-1 mRNA的变化规律,为研究禽类产蛋机制提供基础数据。     

雌性哺乳动物促卵泡素受体与促黄体素受体的结构与表达

本文从雌性哺乳动物促卵泡素受体(FSHR)和促黄体素受体(LHR)的结构及在卵泡发育过程中的表达和功能加以综述,指出FSHR及LHR在卵泡的生长发育、优势化及排卵等方面具有重要的功能。     

不同培养时间对鸡卵泡颗粒细胞孕酮、雌激素分泌水平及FSHR、LHR基因表达的影响

目的：研究培养不同时间鸡卵泡颗粒细胞孕酮和雌激素的分泌水平,促卵泡素受体（FSHR）和促黄体素受体（LHR）的基因表达水平,推断体外培养时间对颗粒细胞激素分泌及相关受体基因表达的影响。方法：通过细胞体外培养的方法,分别于0 h、24 h、48 h、72 h、96 h收集鸡卵泡颗粒细胞上清液,采用ELISA法测定细胞上清液内的孕酮及雌激素分泌水平,并采用荧光定量PCR技术检测颗粒细胞内FSHR和LHR基因表达情况。结果：在培养初期0 h~48 h孕酮和雌激素分泌量显著降低（P < 0.05）,随着培养时间增加到72 h两种激素的分泌量又开始增加,并达到培养初期水平,培养至96 h细胞内孕酮和雌激素分泌量再次降低;颗粒细胞FSHR和LHR mRNA的表达水平则随着培养时间的增加而降低（P < 0.05）。结论：体外培养的卵泡颗粒细胞内孕酮和雌激素的分泌量随体外培养时间的延长呈先降低后升高的趋势,可能与体外培养细胞的生长状态相关,从整体上看随着培养时间的延长,细胞内孕酮和雌激素的分泌量均降低,可能与两种促性腺激素受体FSHR和LHR基因表达量下降相关。     

猪下丘脑和垂体中生长激素受体、胰岛素样生长因子1型受体的发育性变化

GH和IGF-1可作用于垂体或/和下丘脑负反馈性地调节垂体GH的分泌,而这种负反馈作用必须通过下丘脑或垂体的GHR和IGF-1R来实现.为研究猪垂体GH分泌负反馈调节的发育性变化和品种特点,分别在0、3、20、30、90、120和180日龄随机选取纯种雄性二花脸猪和大白猪各4头,屠宰并取下丘脑及垂体,用相对定量RT-PCR分析下丘脑和垂体GHR和IGF-1R mRNA水平.结果表明下丘脑GHR mRNA表达呈明显的时序性变化,在0到120日龄期间呈逐渐上升趋势,180日龄时显著下降(P＜0.05),提示在快速生长期,GH负反馈调控机制逐渐加强.下丘脑GHR mRNA表达还表现明显的品种间差异,在0到180日龄期间大白猪均显著高于二花脸猪(P＜0.05);而垂体GHR mRNA表达相对稳定,品种和年龄间差异不显著,提示GH的负反馈作用位点可能主要在下丘脑.IGF-1R与GHR的表达发育模式不同.下丘脑IGF-IR mRNA的表达相对稳定,无显著的年龄、品种间差异;而在垂体,大白猪和二花脸猪IGF-1R mRNA水平在出生时均较高,随后显著下降(P＜0.05),20日龄后逐渐上升至90日龄的较高水平,随后再次下降.大白猪垂体IGF-1 mRNA表达在30日龄和90日龄时显著高于二花脸猪(P＜0.05),而180日龄时二花脸猪垂体IGF-1R mRNA水平却显著高于大白猪(P＜0.05).结果提示,IGF-1长环负反馈作用位点可能不在下丘脑,而主要在垂体.     

IGF-Ⅰ及其受体、IGF结合蛋白-2和LH受体mRNA在卵泡中的表达

利用原位杂交和原位DNA－3’末端标记的方法研究了胰岛素样生长因子河（IG－I）、IGF－I受体、IGF结合蛋白－2、和促性腺激素受体的信使核糖核酸（mRNA）在不同生长与闭锁阶段的大鼠卵巢卵泡中表达的变化。结果表明：IGF－I主要在正常生长的初级卵泡、窦前卵泡和小窦状卵泡中表达。在各生长与成熟阶段的卵泡中都检测到IGF－I受体mRNA,闭锁卵泡的IGF－I受体表达降低。窦前与窦状的生长和闭锁卵泡均表达IGFBP－2。促卵泡激素（FSH）受体在窦前和小窦状卵泡的表达水平比其在大卵泡中的高。窦前与小窦状卵泡仅在膜细胞中表达黄体生成素（LH）受体mRNA,大卵泡的膜细胞与颗粒细胞均表达LH受体,在闭锁卵泡中仅在膜细胞中观察到LH受体的信号。综上结果,提示IGF－I,IGF－I受体和FSH受体在窦前和小窦状卵泡中的协同表达对卵泡的早期发育有重要作用。LH受体mRNA特异地在大卵泡的颗粒细胞中表达可能与优势卵泡选择相关。     

胰岛素样生长因子-Ⅰ(IGF-Ⅰ)、IGF结合蛋白-2和LH受体mRNA在卵泡闭锁过程中的表达及调节

研究了IGF-Ⅰ、IGF结合蛋白-2(IGFBP-2)和促黄体激素受体(LHR)mRNA在卵泡闭锁过程中的表达及调节.给26日龄大鼠注射15 IU PMSG,经检测,证实PMSG处理48 h后,一些小窦状卵泡的颗粒细胞已发生凋亡;96 h在排卵前卵泡中已可检测到凋亡细胞;120 h大多数的排卵前卵泡中均出现大量的凋亡细胞.48～120 h IGF-Ⅰ主要在窦前卵泡和小窦状卵泡表达;48与96 h,窦前与窦状卵泡的膜细胞均表达高水平的IGFBP-2.在48 h,颗粒细胞中有LHR的强信号,但在96和120 h,颗粒细胞的LHR表达减弱(P＜0.001).表皮生长因子(EGF)和IGF-Ⅰ均抑制窦前和窦状卵泡颗粒细胞凋亡.同时观察到EGF促进IGF-Ⅰ mRNA表达,IGF-Ⅰ刺激排卵前卵泡表达LHR mRNA.上述结果表明,各级卵泡的闭锁可能均受EGF和IGF-Ⅰ相互作用的调节.     

鸭各级卵泡中抑制素α和抑制素／活化素βA亚基信使RNA表达丰度的研究

采用非常灵敏的定量竞争RT朠CR技术对鸭各级卵泡中抑制素π和ρA亚基的mRNA表达丰度作了研究,发现在各级卵泡中均有此两亚基mRNA的表达,大卵泡中的π亚基表达量要高于ρA亚基.π亚基mRNA在小黄卵泡(SYF)中表达量最高,以其mRNA的平均相对丰度为1.00,则F1、F2、F3、F4/5、LWF(大白卵泡)中π亚基mRNA的平均相对含量分别为0.26±0.05、0.28±0.07、0.57±0.12、0.98±0.09、0.026±0.006.ρA亚基mRNA在F1中表达量最高,以其mRNA的平均相对丰度为1.00,则F2、F3、F4/5、SYF、LWF中ρA亚基mRNA的平均相对含量分别为0.218±0.09、0.111±0.03、0.058±0.011、0.053±0.013、0.005±0.002.结果显示,随着卵泡的增大π亚基表达量降低,ρA亚基却增加,说明在卵泡发育过程中,π和ρA亚基的表达是独立调节的,另外,ρA亚基在F1中的大量表达,说明F1可能是形成二聚体抑制素/活化素的主要场所.     

鸭卵泡及精巢中卵泡抑素和抑制素／活化素βB亚基mRNA表达的研究

卵泡抑素是与抑制素/活化素功能密切相关的一种糖蛋白激素,采用定量竞争RT-PCR技术对鸭各级卵泡及成熟与未成熟精巢中的卵泡抑素和抑制素/活化素βB亚基的mRNA表达丰度进行了研究,发现在上述组织中均有此两基因mRNA的表达,且只在小卵泡中表达较丰富.卵泡抑素在小黄卵泡(SYF)中表达量最高,以其mRNA的平均相对丰度为1.00,则F1(最大卵泡)、F2、F3、F4、F5、F6～8,LWF(大白卵泡)、TI(未成熟精巢)和TM(成熟精巢)中卵泡抑素mRNA的平均相对含量分别为0.0ll±0.004、0.019±0.006、0.02l±0.009、0.028±0.007、0.075±0.023、0.15±0.072、0.29±0.068、0.037±0.0l1和0.012±0.004.βB亚基也在小黄卵泡中表达量最高,以其mRNA的平均相对丰度为1.00,则F1、F2、F3、F4、F5、F6～8、LWF、TI和TM中B亚基mRNA的平均相对含量分别为0.009±0.003、0.013±0.005、0.019±0.007、0.023±0.006、0.29±0.084、0.84±0.093、0.031±0.008、0.38±0.072和0.046±0.013.结果显示卵泡抑素和βB亚基mRNA的表达模式是相似的,二者在小卵泡中均出现大量表达说明活化素B(βB-βB)的生物活性可能受卵泡抑素的紧密调节,二者共同在卵泡早期发育中起重要作用.     

生长激素受体的研究进展

生长激素（GH）在促进动物生长、发育等代谢过程中起着重要作用,GH发挥生理作用的第一步是与靶细胞膜表面的生长激素受体（CHR）结合。现已基本阐明了CHR的结构及由CHR介导的信号转导途径,对GHR基因表达调节的机制也有了一定的了解。GHR是由约620个氨基酸组成的单链跨膜糖蛋白,其胞外区、跨膜区及胞区内分别由约245、25及350个氨基酸组成。由GHR介导的信号转导途径主要有：①酪氨酸激酶系统;②蛋白激酶C途径;③胰岛素受体底物途径。营养状况及GH等内分泌因子对GHR的表达也有调节作用。     

胰岛素样生长因子-Ⅰ受体和LH受体mRNA在黄体中的表达及调节

研究了大鼠卵巢黄体中胰岛素样生长因子-Ⅰ受体(IGF-ⅠR)与促黄体激素受体(LHR)的表达关系及调节. 在动情周期中, LHR阳性黄体能表达高水平的IGF-ⅠR mRNA. 动情期黄体的LHR表达水平降低, 而IGF-ⅠR表达仍较高. 假孕第1天黄体中LHR表达较低,第8天达最高水平, 第14天表达已降低到检测不到的程度. 与LHR表达不同, 在第1天假孕黄体中IGF-ⅠR的表达已较高, 此后至第14天在黄体中均检测到IGF-ⅠR信号. 孕鼠黄体同时表达LHR 和IGF-ⅠR mRNA. IGF-Ⅰ刺激LHR mRNA在黄体中的表达. 前列腺素(PG)F抑制IGF-ⅠR和LHR的表达. PGE2能消除PGF对IGF-ⅠR和LHR mRNA表达的抑制作用. 上述结果提示:IGF-Ⅰ可能通过其受体的表达变化参与调节黄体功能、维持黄体结构. PGF抑制IGF-ⅠR的表达可能是黄体萎缩的机制之一.     

陕西省森林群落乔灌草叶片和凋落物C、N、P生态化学计量特征

研究陕西省121个森林群落中乔叶、灌叶、草叶和凋落物4个层次的C、N、P化学计量学特征及其变异性,分析了各层次间及其与地理因子间的关系.结果表明： 乔叶C、N含量最高,草叶P含量最高,凋落物C、N、P含量均最低.乔叶、灌叶、草叶和凋落物的C∶N∶P分别为439.4∶14.2∶1、599.2∶13.5∶1、416.5∶13.3∶1、504.8∶15.5∶1,乔叶、灌叶、草叶N∶P值差异不显著,但均与凋落物N∶P值差异显著,表明不同生活型活体植物叶片的N∶P值具有一定的稳定性.与N含量相比,C和P含量在各层次间表现出更好的相关性;C、N、P含量及其比值在草叶与乔叶之间较在灌叶与乔叶之间表现出更好的相关性;凋落物与乔叶和草叶之间C、N、P含量均表现出显著正相关,与灌叶只在P含量之间存在显著正相关.灌叶N、P含量随纬度增加而增加,草叶N、P含量随经度增加而降低,海拔对叶片和凋落物C、N、P含量及其比值的影响较小,只有乔叶N∶P与海拔之间存在显著负相关.各地理因子对草叶的影响大小表现为经度>纬度>海拔,对乔叶、灌叶和凋落物的影响大小表现为纬度>经度>海拔.     

条背萤成虫与幼虫发光器的超微结构观察

对水生萤火虫——条背萤Luciola substriata(Gorham)成虫和幼虫发光器的超微结构进行研究。结果表明,成虫发光器由明显的2层组成:反射层和发光层。反射层由排列紧密的“尿酸囊泡”构成,具有发达的气管结构,对光起反射作用;发光层由大量发光细胞构成,内含典型的发光颗粒、线粒体、内质网及大量糖原,该层通过发光细胞胞质内的生化反应而发光。2层均由非细胞层膜包被,间距25~30μm。发光器腹节由外向内依次为表皮、发光层、反射层和内部细胞层。幼虫发光器球形,由背射层和发光层构成,由非细胞层膜包被。背射层由单层柱状细胞构成,内含大量“尿酸囊泡”。发光层细胞膜相互绞缠,含有2种类型的发光颗粒:“致密”型和“凋亡”型,含有大量的线粒体和无定形颗粒,发光细胞之间分布着大量的气管、微气管及神经末梢,可观察到神经突触。与条背萤相比,陆生种成虫反射层和发光层均无非细胞层膜包被,2层间无明显间距,发光颗粒形状不规则,气管通常形成2分支;陆栖种幼虫发光层形状差异较大,背射层由单层或2~4层细胞构成;相似点在于,成虫发光器都由均由反射层和发光层构成,发光细胞内都含发光颗粒、线粒体及大量糖原,都具有发达的气管结构,发光颗粒相似。幼虫发光器都由背射层和发光层构成,都具有发达的气管和直接的神经支配,发光颗粒相似,都由非细胞层膜包被。     

Recovery of pristine boreal forest floor community after selective removal of understorey, ground and humus layers

In boreal spruce forests that rarely experience extensive disturbances, fine-scale vegetation gaps are important for succession dynamics and species diversity. We examined the community implications of fine-scale gap disturbances by selective removal of vegetation layers in a pristine boreal spruce forest in Northern Finland. The aim was to investigate how the speed of recovery depends on the type of disturbance and the species growth form. We also wanted to know if there appeared changes in species composition after disturbance. Five different treatments were applied in the study: Control, removal of the ground layer (bryophytes and lichens), removal of the understorey layer (dwarf shrubs, herbs and graminoids), removal of both the ground and understorey layers, and complete removal of the vegetation and humus layers above the mineral soil. The vegetation recovery was monitored in terms of cover and species numbers over a 5-year period. Understorey layer cover, composed mainly of clonal dwarf shrubs, recovered completely in 4 years in treatments where the humus layer remained intact, whereas ground layer cover did not reach the control level in plots from where bryophytes and lichens were removed. Recovery was faster in terms of species number than species cover. Bryophytes, graminoids and dominant dwarf shrubs appeared in all disturbed plots quickly after disturbance. Seedlings of trees appeared exclusively in disturbed plots. Graminoids dominated after the removal of humus layer. The results indicate that the regeneration of forest floor after small gap disturbance occurs mainly by re-establishment of the dominant species. Although destruction of the humus layer leaves a long-lasting scar to the forest floor, exposing of mineral soil may enhance the sexual reproduction of dominant species and the colonization of weaker competitors.     

古尔班通古特沙漠南缘不同林龄人工梭梭林主林层和更新层特征

对古尔班通古特沙漠南缘6个不同林龄人工梭梭种群的结构特征、天然更新及影响因子进行研究.结果表明: 7年生人工梭梭林能天然更新,17年生时幼苗(高度H＜30 cm)、幼树(30≤H＜50 cm)和小树(H≥50 cm)的密度均达到“优”级,小树的平均高度和基径为1.10 m和1.91 cm.20年生人工梭梭林内母树长势最好,35%的个体树高在2.50～3.00 m,23.1%个体的基径达8.00～10.00 cm; 33年生人工梭梭林内母树高和径生长明显衰退,但自然落种更新能力依然较强; 梭梭自然落种更新密度与0～100 cm土层土壤有效氮含量的关联度最大(0.87),其次是土壤速效磷(0.84)及土壤水分含量(0.79),pH 8.1～8.6的土壤未限制更新层的营养生长.在主林层,与整个更新层个体密度关联性最高的是人工梭梭林母树生物量(0.77),而与更新层小树密度关联性最高的是种植密度(0.71)和母树年龄(0.70).     

宁夏罗山自然保护区3种典型林分凋落物和土壤层水源涵养能力综合评估

森林凋落物层和土壤层是森林生态水文效应中的主要贡献层，对森林生态系统水土保持功能和水源涵养能力有重要影响。对比宁夏罗山自然保护区3种典型林分类型凋落物和土壤层水文效应的变化规律和水源涵养能力大小，为该地区的森林生态水文、水土保持和森林管理提供科学依据。以该自然保护区青海云杉纯林、油松纯林和青海云杉油松混交林为研究对象，运用称量、室内浸泡、环刀法和回归分析法对凋落物和土壤层的水文效应进行测定和拟合，并使用熵权法对二者的水源涵养能力进行评估。结果表明：（1）青海云杉油松混交林凋落物的总厚度和总储量显著高于青海云杉纯林和油松纯林（P<0.05），3种林分类型的半分解层的厚度和储量高于未分解层。（2）凋落物层最大持水量范围为63.29-95.08t/hm²，最大持水率范围为335.97%-353.85%，有效拦蓄量范围为34.09-63.92t/hm²，三者均为青海云杉油松混交林>云杉纯林>油松纯林。（3）3种林分类型的凋落物持水量（Q）与浸泡时间（t）呈对数函数关系，吸水速率（V）与浸泡时间（t）呈幂函数关系。（4）从3种林分类型的土壤物理性质和持水特性得出，3种林分类型的土壤层水文效应的等级排序为云杉油松混交林>油松纯林>青海云杉纯林。（5）凋落物层和土壤层的水源涵养能力大小为青海云杉油松混交林（0.43）>油松林（0.3）>青海云杉林（0.27）。综合来看，青海云杉油松混交林的凋落物层和土壤层的水源涵养能力最优，其次是纯林，说明混交林在水土保持和水源涵养方面比纯林更具优势。     

Structure of the adenohypophysis in juvenile harp seal,Pagophilus groenlandicus

Summary A study was made of the morphology of the adenohypophysis in immature harp seals and the fine structure of cellular components of the pars tuberalis, pars intermedia and pars distalis was described.The pars intermedia composed 8–15% of the hypophysis and contained colloid filled vesicles similar to those found in the other mammalian species.The pars distalis cells were grouped into more or less well defined regions, thus facilitating the correlation of cellular identification from both light and electron micrographs. Five chromophilic cell types were tentatively identified, one acidophil (putative somatotroph), four basophils (3 putative gonadotrophs and one cell type with the characteristics of both corticotrophs and thyrotrophs) and non-granulated stellate cells. The absence of a positive prolactin cell identification was thought to be due to the immaturity of the seals used in the study.The mercury exposure experiment was supported by a contract grant from the Halifax Laboratory of Fisheries and Marine Service. Drs. Uthe and Freeman of that laboratory also carried out the methyl mercury analyses. We recognize the support in maintaining the seals provided by Mrs. C. Rae, Mrs. H. Pedersen, Mr. S. Tessaro and Dr. J. M. Terhune. We also wish to thank Mrs. L. Lin for her technical assistance. Further financial support was provided through operating and development grants. The paper is number 134 in the physiology of migration series     

Internal architecture, origin-insertion site, and mass of jaw muscles in Old World hamsters

The jaw muscle (i.e., masticatory, suprahyoid, and extrinsic tongue) anatomy and mass were examined in four genera of Old World hamsters (cricetine murids), Mesocricetus, Cricetulus, Tscherskia, and Phodopus. The masseter was the largest and most complicated of the muscles examined. In the superficial layer, a few ventral fibers form a small medially turned portion with an insertion site more similar to those of sciurids than of other murids. In Mesocricetus, the superficial layer has a discrete anteroventral portion that has not been reported for other murid rodents. Examination of the fiber attachment sites indicated that the deep layer contains four parts and the medial layer contains three parts. The deep layer originates from two aponeuroses that are firmly connected to each other at their anterior ends and lie along the zygomatic arch. The aponeurosis of insertion for the deep layer is situated along the masseteric ridge and the dorsal border of the angular process, but is absent in its middle part, consistent with reports in two relatives, sigmodontine and arvicoline murids. In cricetine murids, unlike in other rodents, fibers insert on the dorsal narrow strip of the posterior mandibular aponeurosis, not on its broad medial aspect. The relative mass of some masticatory and suprahyoid muscles is related to body mass. Small species (Cricetulus and Phodopus) have relatively larger masseter and mylohyoid muscles and smaller temporalis and geniohyoid muscles than large species (Mesocricetus and Tscherskia).     

Aggregation Behaviors of Glutens,Glutenins and Gliadins from Various Wheats

The aggregation behavior was investigated by the method presented in the previous paper, with the glutens separated from ten kinds of wheat flours different in the mixing property. It was shown that the rate constant of aggregation, k, increased in the order of the weak, medium and strong flour. This is an additional evidence for the correlation between the aggregation behavior of the separated gluten and the mixing property of the flour, which was suggested in the previous paper with four kinds of flours. The aggregation behaviors of glutenin and gliadin were also investigated after fractionation of gluten. A good correlation was observed between the τ₁₀/C values of gluten and glutenin separated from it, showing that the difference of the aggregation behavior of gluten is mainly due to the nature of glutenin.     

Single layer colloid centrifugation technique improves motility,viability and chromatin integrity of ram spermatozoa after thawing

The cell membrane of ram spermatozoa is more sensitive to the freezing process than in other species due to its composition. As a result, the quality and viability of frozen thawed ram spermatozoa are often poor, which together with the specific structure of the ewe's cervix are the main reasons for lower fertility in ewes after intracervical insemination. In the present study we investigated the effects of semen centrifugation through a single layer of a species-specific colloid (Androcoll-O) on post-thaw quality of ram spermatozoa. Motility, viability and morphology were analysed 0, 6, 12 and 24 h after thawing. DNA fragmentation index (%DFI) of the samples was assessed 0 h after thawing, by SCSA™. Membrane and acrosome integrity of spermatozoa were analysed by Sybr-14/PI/PNA test 0 h after thawing.The proportion of motile spermatozoa was significantly higher in SLC – selected samples in comparison to control (not SLC – selected) samples at 0, 6, 12 (P < 0.001) and 24 h (P < 0.05). The proportion of viable spermatozoa was also significantly higher in SLC - selected samples in comparison to control samples at all times (P < 0.001). The proportion of abnormal acrosomes and morphologically abnormal spermatozoa (MAS) were significantly lower in SLC – selected samples compared to control samples at all times (P < 0.001). Analysis of chromatin stability revealed significantly lower %DFI values in SLC – selected samples compared to control samples (P < 0.001). The SYBR-14/PI/PNA test also revealed significantly better values in SLC – selected compared to control samples (P < 0.05). In conclusion, single layer colloid centrifugation significantly improved post-thaw quality and longevity of ram spermatozoa, making it suitable for artificial insemination initiatives.     

鸡胚胎干细胞的分离、培养和鉴定

SNL cells (permanent line of irradiated mouse fibroblast cells), primary mice embryonic fibroblasts (PMEF) cells and primary chicken embryonic fibroblasts (PCEF) cells were respectively used as the feeder cells for chicken embryonic stem cell culture. The isolated blastoderm cells front the stage X embryos of chicken were cultured in Dulercco‘‘ s Modified Eagle Medium (DMEM) supplemented with leukemia inhibitory factor (LIF, 1 000 IU/ml), basic fibroblast growth factor (bFGF 10 ng/ml) and stem cell factor (SCF, 5 ng/ml). The alkaline phosphatase (AKP) test, differentiation experiment in vitro and chimeric chicken production were carried out. The resuts showed that culture on feeder layer of PMEF yielded high quality CES cell colonies. The shape of typical CES clone showed as follows: nested aggregation (clone) with clear edge and round surface as well as close arrangement within the clone. Strong positive AKP reactive cellswere observed. On the other hand, the fourth passage CES cells could differentiate into various cells in the absence of feeder layer cells and LIF in vitro. The third and fourth passage cells were injected into the subgerminal cavity of recipient embryos at stage X. The manipulated embryos were incubated until hatching. Of 269 Hailan embryos injected with CES cells of Shouguang Chickens, 8.2 % (22/269) survived to hatching, 3 feather chimeras had been produced, which suggests that an effective culture systems were established and it could promote the growth of CES cells and maintain them in an undifferentiated state .