无菌培养液

In vitro, we cultured human and rabbit retinal Müller cells by way of mixed enzyme digestion and tissue adherence, using human retina of an abortus of 6 months and healthy rabbit.

取孕6个月引产胎儿，死后2h内无菌条件下摘取眼球，剥取视网膜组织，混合酶消化法培养人视网膜Müller细胞；取健康新西兰大白兔，死后以最快的速度无菌条件下摘取眼球，组织块贴壁法培养兔视网膜Müller细胞；利用含胎牛血清的高糖DMEM作为培养液，不添加生长因子。

The diluted monoplast suspension was equally assigned into 3 samples, and separately used for simple modified differential adherence, simple serum-free conditioned medium, and modified differential adherence ＋ serum-free conditioned medium.

①无菌条件取大鼠嗅球组织，消化、离心去除上清液后，用含体积分数为0.2胎牛血清的DMEM/F-12培养基重悬，稀释的单细胞混悬液均分成3份，分别用于单纯改良差速贴壁、单纯无血清条件培养液纯化和改良差速贴壁＋无血清条件培养液纯化的实验。

METHODS: Olfactory bulb was sterilely collected from rats, digested, and centrifuged. After removal of the supernatant, the olfactory bulb was resuspended with DMEM/F-12 medium containing 0.2 volume fraction of fetal bovine serum. The diluted monoplast suspension was equally assigned into 3 samples, and separately used for simple modified differential adherence, simple serum-free conditioned medium, and modified differential adherence + serum-free conditioned medium.

①无菌条件取大鼠嗅球组织，消化、离心去除上清液后，用含体积分数为0.2胎牛血清的DMEM/F-12培养基重悬，稀释的单细胞混悬液均分成3份，分别用于单纯改良差速贴壁、单纯无血清条件培养液纯化和改良差速贴壁+无血清条件培养液纯化的实验。

To compare the growth characteristics of non-hematopoietic adult stem cells derived from rat fetal blood and rat bone marrow in vitro, and to study the differentiation of these stem cells into neuron-like cells in vitro,the fetal blood of pregnant rats and bone marrow of adult rats were sterilely collected; mononuclear cells were isolated by using standard Ficoll-hypague techniques and then cultured in DMEM/LG containing 10% fetal bovine serum.

为了比较大鼠胎血和骨髓中非造血成体干细胞（non-hematopoietic adult stem cells，NASC）体外培养过程中的生长特性及体外诱导两者向神经元样细胞分化的异同，在无菌条件下采集孕大鼠的胎血和成年大鼠的骨髓，用Ficoll-hypague分离液分离出单个核细胞后，种植于含10%胎牛血清的DMEM/LG培养液内。收获的NASC传代培养，用流式细胞仪检测细胞的免疫表型。

To compare the growth characteristics of non-hematopoietic adult stem cells derived from rat fetal blood and rat bone marrow in vitro, and to study the differentiation of these stem cells into neuron-like cells in vitro,the fetal blood of pregnant rats and bone marrow of adult rats were sterilely collected; mononuclear cells were isolated by using standard Ficoll-hypague techniques and then cultured in DMEM/LG containing 10% fetal bovine serum.

为了比较大鼠胎血和骨髓中非造血成体干细胞（non-hematopoietic adult stem cells，NASC）体外培养过程中的生长特性及体外诱导两者向神经元样细胞分化的异同，在无菌条件下采集孕大鼠的胎血和成年大鼠的骨髓，用Ficoll-hypague分离液分离出单个核细胞后，种植于含10%胎牛血清的DMEM/LG培养液内。

To compare the growth characteristics of non-hematopoietic adult stem cells derived from rat fetal blood and rat bone marrow in vitro, and to study the differentiation of these stem cells into neuron-like cells in vitro,the fetal blood of pregnant rats and bone marrow of adult rats were sterilely collected; mononuclear cells were isolated by using standard Ficoll-hypague techniques and then cultured in DMEM/LG containing 10% fetal bovine serum. The acquired NASCs were subcultured for passage.

为了比较大鼠胎血和骨髓中非造血成体干细胞（non-hematopoietic adult stem cells，NASC）体外培养过程中的生长特性及体外诱导两者向神经元样细胞分化的异同，在无菌条件下采集孕大鼠的胎血和成年大鼠的骨髓，用Ficoll-hypague分离液分离出单个核细胞后，种植于含10%胎牛血清的DMEM/LG培养液内。

And try to find out the effect of TSA on the proliferation and differentiation of rat tracheal stem cell. Methods 1.Preparation of 5-Fu Model and TSA treatment Two weeks" Wistar rats provided by the animal center of the China Medical University. The animals were sacrificed by 10% Chloral Hydrate 0.4ml/100g. Then the tracheas were excised aseptically. Then they were douched with PBS and cultured in 1:1 mix of Dulbecco"s modified Eagle"s medium and Ham"s F-12 medium (DMEM/F12) containing 120 mg/ml 5-Fu and 10% fetal bovine serum for 12h at 37°C. The tissues were then refreshed by culturing in DMEM/F12 containing 10% FBS and 200nM TSA.

1、制备气管损伤模型及剥离上皮取约200克左右的Wistar大鼠，雌雄不限(中国医科大学实验动物中心提供)，腹腔注射10%水合氯醛0.4ml/100g，无菌条件下取出气管，无菌PBS反复冲洗，洗净血液和粘液，置于DMEM/F12培养液中(含10%胎牛血清)，于培养液中加入终浓度为100mg/ml的5-Fu，37℃，5%CO2孵育12小时，弃去上述培养液，换成新鲜DMEM/F12液(含10%胎牛血清)，同时加入终浓度为200nM的TSA继续培养，于换液后3、6、12、24、48小时分别10%中性福尔马林固定，石蜡包埋，制成4μm厚的组织切片。

The ELISA assay established with the crude antigen-specfic monoclonal antibodies could detect both of the clinical and environmental isolates of Aspergllius, while the other assay could only detect Aspergillus fumigatus of both clinical and environmental isolates.And no cross reaction with the cell culture of Penialllium marneffei and Candidas was observed with the two methods.

用mAbs-1建立的双抗体夹心ELISA法可检测19种常见曲霉株培养液；用特异性针对烟曲霉抗原单克降抗体(mAbs-2)建立的双抗体夹心ELISA法可特异性检测临床和环境分离株烟曲霉培养液；与其他曲霉株无交叉反应；2种双抗体夹心ELISA法与马尔尼菲氏青霉菌及念珠菌培养液均无交叉反应。

objective to establish immunological methods specific for detecting antigens in different groups of monoclonal antibodies.methods indirect immnofluorescence assay was applied to identify specificity of the two groups of monoclonal antibodies prepared with crude antigen and recombinant antigen of aspergillus fumigatus,respectively.two different double monoclonal antibody sandwich elisa assays established with the two groups of antibodies were performed to detect antigents in the cell culture supermatants of 19 common species of aspergillus,penicillium marneffei,and 5 species of candidas.results the results of indirect immnofluorescence assay indicated that the monoclonal antibodies prepared with crude antigen of aspergillus fumigatus were specific for antigens in both clinical isolates and environmental isolates of aspergillus, whereas the other group of monoclonal antibodies was proved to be specific for aspergillus fumigatus of both clinical and environmental isolates.the elisa assay established with the crude antigen-specfic monoclonal antibodies could detect both of the clinical and environmental isolates of aspergllius, while the other assay could only detect aspergillus fumigatus of both clinical and environmental isolates.and no cross reaction with the cell culture of penialllium marneffei and candidas was observed with the two methods.conclusion the elisa assays can detect both of the clinical and environmental isolates of aspergillus,and differentiate aspergillus fumigatus from other species of aspergillus.

目的 用2组曲霉单克隆抗体建立特异性识别不同种类曲霉抗原的检测方法。方法采用天然烟曲霉抗原免疫，获得广谱针对曲霉抗原的单克隆抗体；采用重组烟曲霉抗原获得特异性针对烟曲霉抗原的单克隆抗体，用间接免疫荧光鉴定，并分别建立2种双抗体夹心elisa法，对19种常见的环境和临床分离曲霉株、马尔尼菲氏青霉菌及念珠菌培养液进行检测。结果间接免疫荧光显示，用天然烟曲霉抗原免疫获得的单克隆抗体(mabs-1)可广谱识别多种曲霉分离株，而重组烟曲霉抗原获得的单克降抗体(mabs-2)仅能特异性结合临床和环境分离的烟曲霉抗原。用mabs-1建立的双抗体夹心elisa法可检测19种常见曲霉株培养液；用特异性针对烟曲霉抗原单克降抗体(mabs-2)建立的双抗体夹心elisa法可特异性检测临床和环境分离株烟曲霉培养液；与其他曲霉株无交叉反应；2种双抗体夹心elisa法与马尔尼菲氏青霉菌及念珠菌培养液均无交叉反应。

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。

And Pharaoh spoke to Joseph, saying, Your father and your brothers have come to you.

47：5 法老对约瑟说，你父亲和你弟兄们到你这里来了。

Additionally, the approximate flattening of surface strip using lines linking midpoints on perpendicular lines between geodesic curves and the unconditional extreme value method are discussed.

提出了用测地线方程、曲面上两点间短程线来计算膜结构曲面测地线的方法，同时，采用测地线间垂线的中点连线和用无约束极值法进行空间条状曲面近似展开的分析。