传代

The invention discloses a connecting city white duck blastodisk fiber cell system with connecting city white duck embryo as material with preservation number at CGMCC No.1877 in cellular biology domain, which is characterized by the following: the fiber cell does not possess epithelial cell with high purity; the quality of freezing cell is stable; the active ratio can reach between 93.5% and 96.8%; the passage growth is stable and fit for big scale culture.

本发明利用连城白鸭胚胎作为材料，进行初代培养、传代培养及细胞冻存等研究。最终获得高活率、高纯度的连城白鸭胚成纤维细胞系，其保藏编号为CGMCC No.1877属于细胞生物学领域。本发明培养的成纤维细胞无上皮细胞等，细胞纯度高；冻存后细胞质量稳定，活率可达到并维持在93.5％～96.8％之间，传代生长稳定，适合大规模培养。

The colony of embryonic stem cell separated from blastula could gain five strains of embryonic stem cell and go down to posterity F7 in vitro.

从培养的囊胚中分离获得的类胚胎干细胞集落，经传代培养获得了5株胚胎干细胞株，在体外可传代培养到7代，冷冻保存复苏后，干细胞仍能存活、增殖。

The incidence of blue and green mold of citrus was reduced by 25.56% and 10.00% in vivo experiment after 7 days respectively.

生长动态测定和传代试验表明，该菌株生长特性优于出发菌株，连续传代10代，该菌株没有出现退化、回复突变等，在遗传上是稳定的。

Silk gland cell, by the Grace nutrient fluid in vitro culture, then after 150d for primary culture, passage was taken, later each approximately 15d take a passage, after that, the time of passage cut to 10 day.

经210d左右的原代培养后进行传代培养，以后每隔约15d传代一次，后传代时间缩短到10天，当传代时间稳定在7天时细胞开始趋于稳定，此时鉴定细胞系的生物学特性。

We examined RPE by microscopic observation and immmunohistochemical analysis with antibodies against cytokeratin, an epithelial marker, and α-smooth muscle actin, a marker of myoid differentiation.

自成人供体眼获取RPE培养传代，显微镜下观察原代RPE的形态以及传代(3、9)RPE的去分化改变，免疫组化法(角蛋白抗体和平滑肌肌动蛋白抗体)检测传代后RPE免疫标记的改变。

Objective: To determine whether the cells that we obtained from the in vitro culture of human embyonic genital ridges and dorsal mesenteries were the undifferentiated human embryonic germ cells , to find out the best kind of passaging method , and finally to identify these serial passaged cells .

目的：鉴定组织块培养法体外培养人胚胎生殖腺嵴和背侧肠系膜所获得的细胞为未分化的人胚胎生殖细胞；尝试不同时间和不同方法对人EG细胞进行传代培养，探讨最佳传代时机和传代方法；并确定传代培养的细胞仍为未分化的人EG细胞。

Methods Primary human embryonic germ cells were obtained by tissue culturing of embryonic genital ridges and mesenteries from the embryos aged 8 weeks, and then were passaged on the 8th and 16th day using pick-up and digestive passage method. Three kinds of digestive juice were used.

取8周人胚胎的生殖腺嵴和肠背系膜，进行组织块体外培养人胚胎生殖细胞，选择原代培养第8天和第16天的细胞，采用挑取传代法和消化传代法，其中的消化传代法采用三种不同的消化液进行传代培养，记录各代培养的细胞集落数。

Immunohistochemical identification of cultured conjunctival cellsWhen the second cultured generation of conjunctival cells close to confluence, the glass coverslips were removed in situ fixation by methanol and DAB color immunohistochemical staining by CK13 monoclonal antibody in time.3. growth curve of conjunctival cellsThe 1st,3rd,5th passage cells were selectecd randomly and subcultured at the density of 5×10~4/ml,after culturing for another 1-9 days, were counted and compared the cell number every day.

采用消化后组织块法培养结膜原代细胞，并进行传代。2、培养结膜细胞的免疫组化鉴定把第2代的结膜细胞接种于置在培养皿中的盖玻片上，当细胞接近融合时，及时取出作原位甲醇固定，CK13单抗、DAB显色免疫组化染色。3、结膜细胞生长曲线的测定随机选取第1、3、5传代细胞，以5×10~4／ml密度传代、分别培养1-9天，每天分别细胞计数并进行分析比较，重复3次。

Greater than 40 serial MSC passages and 100 cell population doublings have been recorded for some MSC batches. Early and late passage MSCs were defined here as those cultures receiving less than 5 trypsin passages and more than 15 trypsin passages, respectively.

本实验研究了传代超过40代的MSCs和100个细胞集落，并将传代少于5次和多于15次的分别命名为传代早期和晚期的MSCs。

The hFCECs were cultured by sticking tissues piece method that the cornea were divided into the limbus and central tissue piece and digestive method,respectively.For digestive method,the digestive juice of 5mg/ml DispaseⅡ+0.25%trypsin was chosed.D/F12 with 10%fetal bovine serum and 100 U/ml penicillin and streptomycin were used in this study,the culture condition was 37℃and 5%CO2,and culture medium changed every three days2、Passage of hFCECsAfter more than 80%confluenced,cells from corneal limbus hFCECs were digested for 5-15min with different concentrations of trypsin/EDTA at 37℃,and then passaged at a ratio of 1:2.The cells were subcultured on empty plates,mouse 3T3 fibroblast feeder layer,fetal corneal stromal cell feeder l ayer or HTK feeder layer respectively.And D/F12, mouse 3T3 fibroblast conditioned medium,fetal corneal stromal cells conditioned medium or HTK conditioned medium with 10%FBS and 100 U/ml penicillin and streptomycin were used respectively as the culture medium.And the culture medium were changed every three days as well.

方法一、人胎儿角膜上皮原代和传代培养1、原代培养严格无菌操作获取人胎儿角膜片，采用组织块贴壁法、酶消化法原代培养人胎儿角膜上皮细胞。2、传代培养角膜缘部的细胞生长融合达80%以上，不同稀释浓度的胰酶/EDTA消化液消化传代分别接种于空板、小鼠3T3成纤维细胞饲养层、胎儿角膜基质细胞饲养层及HTK饲养层，培养液分别采用D/F12、小鼠3T3成纤维细胞条件培养液、胎儿角膜基质细胞条件培养液、HTK条件培养液。

Since historical times,England ,where the early inhabitants were Celts, has been conquered three times .

从有历史以来，英国，在此地早期居住的是凯尔特人，已经被征服了三次。

Bluetooth OBEX File Transfer Enables the sending and receiving of files on your phone via Bluetooth.

蓝牙OBEX文件移动允许经过蓝牙传送和接受文件。。。。