Newcastle disease virus ( ndv ) strain 695, a thermostable nature avirulent strain, were replicated in embryonated chicken eggsand its rna was extracted from allantoic fluid. referred to the reported sequence of f gene, a pair of primers were designed and synthesized. f gene of ndv b95 strain was amplified by rt - pcr, the pcr products were checked by agrose gel electrophoresis and purified by agrose gel fracion method
利用從國外引進的新城疫熱穩定性天然弱毒b _ （ 95 ）株接種spf雞
胚繁殖病毒，經處理后提取病毒的基因組rna ，參考國內外發表的ndv融合蛋白基因序列，設計一對特異性引物，經反轉錄聚合酶鏈式反應（ rt - pcr ）擴增出約1700bp大小的特異性片段，將此片段回收純化后，利用t - a克隆技術將其克隆到pgem - t - easy克隆載
體中，再轉化大腸桿菌jm109感受態細胞，轉化后經分子量比較、 pcr鑒定和酶切分析篩選陽性克隆。
Under suspension culture without cytokines, the putative eg clonies could be induced to form simple embryoid bodies. after long - time culture, these bodies began to differentiated into various cell types
將消化分散的類eg細胞在無分化抑制因子的類eg培養基中重懸培養， 3 4天後，可見部分細胞團形成簡單的
胚體。
Eg cells of the 2th and 4th passage were akp ( alkaline phosphate ) positive. when cultured on degenerated feeder layers or in suspension, eg c ells formed embryoid bodies ( ebs ) in vitro
當eg細胞脫離飼養層懸浮培養，或在衰老的飼養層上延遲培養時，發現eg細胞或單個存在，或聚集成團，形成類似於早期
胚胎的囊狀
胚體結構
Spontaneous differentiation of es cell d3 line in vitro es cell d3 line via suspension culture can congregate small mass within 24 h, and large numbers of aggregates called embryoid bodies ( ebs ) gradually formed after 2 to 3 days of culture
5 . es一d3細胞系的自主分化es一d3細胞懸浮培養， 24h后可聚集成小的細胞團，第2 ~ 3d時形成大量的類
胚體（ ebs ） 。
There is no different between the fibroblast feeder of mouse and the fibroblast feeder of goat according to blastoysts attached and icm proliferation. es - like cell develop embryoid body - like structure in suspension culture in vitro. es - like cells of goat differentiate into various kinds cell, egg : fibroblast cell, neurolike cell
Es細胞在
體外懸浮培養，形成類
胚體，山羊類es細胞在無飼養層鋪有明膠的皿底上培養，可分化為多種類型細胞，如上皮細胞、成纖維細胞、神經細胞等。
The embryonic shield appeared in 15h, and the fertilized egg entered the middle gastrula stage
15小時出現
胚盾， 15小時55分，
胚體下包一半。
The results of biological tests have demonstrated that allantoic fluid of the first passage virus did n ' t produce macroscopic pathogenic role to chicken embryos and after passaged for four times, gross lesions were observed in chicken embryo. the virus showed typical coronavirus under electron - microscope and it could n ' t form plaque in cef cells and could hemagglutinates chicken red blood cells after treatment with 1 % trypsin. to surprise, the virus replicated in cef cells also showed hemagglutination activity to chicken red blood cells. in addition, the spf chickens which inoculated with the virus isolated from the chicken damaged tissue showed clinical sign and grow lesion, but it ' s gross lesion did n ' t resemble to those of field outbreaks
生物學特性：雞
胚尿囊液經離心、磷鎢酸負染后，電鏡觀察該病毒為典型的冠狀病毒；該毒株的第一代尿囊液對雞
胚無肉眼可見的致病作用，當繼代到第5代后，
胚體嚴重病變；病毒在雞
胚中隨著接種時間的延長，其效價增高， 96h可達到48h的2倍；該毒株可在cef上生長，但不能形成明顯的蝕斑；經1胰酶處理后可凝集雞紅細胞；雞
胚的第四代尿囊液病毒回歸動物
體，病死雞腎臟呈典型的花斑腎，腺胃則未見肉眼可見的病變。
The quality of feeder layer is affected by a lot of factors, such as animal breed, culture medium, passages in vitro and experiment condition, etc. as to the production of feeder layer, there are a few reports about morphological and histologic change when of embryonic body fibroblast when culturing in vitro and cryopreservation, so kunming mouse were chosen as experimental animals and morphological and histologic changes were studied in course of its embryonic body culturing. we expect to offer theoretical foundation to our laboratory for setting up feeder layer storehouse. at the same time, the feasibility of myocardium tissue culturing with fibroblast layer altogether was studied so that established foundation for studied the biological characteristic of heart outside body
小鼠
胚體成纖維細胞的培養是制備飼養層的重要途徑，其制備、傳代及冷凍保存均有不同的研究報道，飼養層的質量受許多因素的影響，如動物的品種、培養液、所傳代數及實驗條件等，關于飼養層制備過程中的
胚體細胞培養、傳代、冷凍后的細胞形態、組織學等方面的研究報道很少，故本實驗以昆明小白鼠為實驗動物，研究其
胚體培養過程中細胞的形態學、組織學等方面的變化，以期為本實驗室建立飼養層細胞庫提供理論依據，同時探討心肌細胞和成纖維細胞層共培養的可行性，以期為心臟生物學特性的
體外研究奠定基礎。
( 2 ) the ratio of nucleus to cytoplasm about fibroblast increased gradually with culturing time in the same generation
（ 2 ）
體外培養的原代小鼠
胚體成纖維細胞隨培養時間的增加，核質比逐漸增加。
Study on the condition of pro - embryoids suspension culture of dendrobium candidum
鐵皮石斛原
胚體液
體懸浮培養條件
Day 4 embryoid body ( 4deb ) cells were derived from es cells and then induced with mbmec - cm into hematopoietic precursor cells. immunocytochemistry staining and flow cytometry were adopted to observe the antigen expressions. rt - pcr method was used to detect the expressions of hematopoietic genes
在誘導小鼠es細胞向造血細胞分化的實驗中，我們採用先將小鼠
胚胎幹細胞形成4天擬
胚體（ day4embryoidbodies ， 4debs ） ，再用骨髓內皮細胞條件培養液誘導4天擬
胚體細胞生成造血干祖細胞。
Embryo almost encircled the yolk - sac in 38h40min. the larva broke out of the egg in 39h50min, at that time, the number of the oil globules decreased greatly
38小時40分
胚體繞卵黃囊約一周， 39小時50分仔魚孵出，仔魚孵出時卵黃囊內的油球已大大減少。
( 4 ) the myocardium cell group on embryonic body fibroblast feeder was cultured successfully. the myocardium tissue grew gradually, its contraction changed with temperature and morphology changed with time, and not all myocardium tissue could contract
（ 4 ）小鼠
胚體成纖維細胞可以與心肌細胞團共培養，並出現自律性收縮，細胞團逐漸增長，其收縮情況隨溫度變化而變化，隨時間變化其形態學發生變化，而且並非所有心肌細胞團都能收縮。
The ebs of different days gained by means of hanging, suspending and suspending following hanging were processed by all - trans retinoic acid ( ra ) for 4 days. during this period, we observed the differentiation process and compared the differentiation ratio of es cells processed by means of hanging, suspending for several days. based on this study, we induced the ebs by ra of different concentration in order to study the effect of ra on the differentiation of es cells into neuron - like cells
首先從
胚體（ ebs ）本身探討es細胞內部調控誘導分化的機制，用懸浮培養、懸滴培養、懸浮轉懸滴培養的方法培養出不同時間的ebs ，再加以ra處理，觀察比較ebs對es細胞神經分化的影響；在此基礎上，用不同濃度的ra處理ebs ，分析誘導劑ra對es細胞神經分化的影響。
1 shape of core plate
1胚體基本形狀
