组织培养

Immunocytochemistry method was used to demonstrate the expression of glial fibrillary acidic protein.

用塑料吸头划刮培养皿，制造机械损伤的模型，继续培养至72 h，分别取出不同时间点的细胞进行胶质细胞增生情况的观察和免疫组织化学染色，对胶质原纤维酸性蛋白表达产物光密度进行图像分析。

METHOD:SD rat aortic tissue was planted with the adherent method of tissue explants. The cellular morphology was observed by inverted phase contrast microscope and the nucleus and cytoplasm were observed after haematoxylin and eosinstaining.

运用组织块贴壁法进行SD大鼠胸主动脉平滑肌细胞培养，并用倒置相差显微镜观察、HE染色后形态学观察以及用免疫组化对培养细胞进行鉴定。

The steps are as the following:1、Preparing the cerebrospinal fluid one hour before the experiment,take some of them for cryopreservation;2、Without paralyse,cuting down the rat head quickly and dislodge the hippocamp part which is used in the experment in freezing liquid,and trim the hippocamp distrct.3、Fixing the hippocamp tissue on the vibratome,to cut down a brain slice with 300μm,use the haustorial tube to move the slice to the preincubate dish,cultivanting it for one hour.Then move it to the record incubation chamfer.4、Preparing the glass micloelectrode and fill it with NaCl,the corcentation is 3mol/L.5、adjusting the recorder,after cultivate the brain slice in the incubation chamfer for 2 hours,the expermentize can be gone on.6、In order to ensure the accurate of the experimental result,use only one medicine concentration for one brain slice in every experiment.

本实验研究方法是：1、实验前1小时配制好所需脑脊液并充以95% O2和5% CO2的混合气，取小部分冷冻保存；2、在乙醚麻醉下，快速断头并在冷冻液里取出包含海马的大脑部位，修整出所需海马区；3、将海马组织固定于振动切片机上切出300μm厚的海马脑片，用广口吸管转移至预孵育皿培养一小时，后再转移到记录用孵育槽内；4、拉制好玻璃微电极，并充以3mol/L的NaCl；5、调试好记录仪器，将脑片在孵育槽内培养2小时后进行实验；6、为了保证实验结果的准确性，每一块脑薄片只进行一个浓度的药品实验。

Methods Autologous cord blood serum was used to culture cord mesenchymal stem cells. An enzyme digesting method was used to isolate mesenchymal stem cells from the cord. Morphology, immunotyping and hepatogenic differentiation of these cells were assayed.

分离自体脐血清用于后续细胞培养，利用酶消化法从脐带组织中分离培养间充质干细胞，检测其形态和免疫表型，并采用两步法诱导其向肝细胞诱导分化。

Part I: Primary culture and purification of skeletal muscle derived satellite cells for tissue engineering applications To investigate separation and purification of skeletal muscle satellite cells with improved incontinuous density Percoll gradient centrifugation technique for tissue engineering applications.

第一部分：种子细胞—肌卫星细胞的原代培养及鉴定本部分通过应用改良Percoll非连续密度梯度离心法分离纯化培养新西兰兔骨骼肌干细胞—肌卫星细胞，探讨为组织工程膀胱的构建提供种子细胞的可行性。

Detect that whether the ADSCs were differentiated inducedly into osteoblasts or adipogenic cells by Alkaline phosphatase staining、Von kossa staining and Oil red "O"staining after the ADSCs were cultured in inductive basic medium containing osteogenic induction agent and adipogenic inducers for four weeks.②.

①。采用密度梯度离心法加贴壁培养法对兔脂肪组织进行了分离纯化，CD44免疫组化法分析ADSCs表面标志；用含成骨、成脂诱导培养基培养4周后，行碱性磷酸酶染色、Von kossa染色及油红&O&染色检查ADSCs是否向成骨、成脂细胞定向分化。②。

Objective: Identify the human embryonic germ cells by observing the expression of its specific surface markers SSEA-1、SSEA-3. Methods: Human embryonic genital ridges, dorsal mesenteries(5-9 weeks post-fertilization) were cultured without any cytokines, while the fibroblasts derived from the isogenous embryo were used as the feeder layer.

目的：组织块培养法培养人胚胎生殖干细胞，通过检测细胞所表达的阶段特异性胚胎抗原1 和3(stage-specific embryonic antigen-1、3 ，SSEA-1、SSEA-3)，对未分化的人EG 细胞进行特异性的生物学鉴定。

To study the molecular mechanism of Larix somatic embryogenesis, a differentially expressed cDNA library of Larix somatic embryo in the period of maturation was constructed using suppression subtractive hybridization. The cDNA from the cultures at the stage of somatic embryo maturation of embryogenic cell line Y35 of L. leptolepis ×L. principis-rupprechtii was used as the tester and the cDNA from its subcultured callus was used as the driver. Eight hundreds randomly selected positive clones were sequenced, and 468 UniGenes were obtained finally.

为研究落叶松体细胞胚胎发生的分子机理，文章以日本落叶松×华北落叶松杂种无性系胚性细胞系Y35体细胞胚成熟阶段培养物的cDNA为实验组，继代培养阶段胚性愈伤组织的cDNA为对照组，利用抑制性消减杂交技术(Suppression subtractive hybridization， SSH)构建了体细胞胚成熟阶段的差异表达基因文库。

To study the molecular mechanism of Larix somatic embryogenesis, a differentially expressed cDNA library of Larix somatic embryo in the period of maturation was constructed using suppression subtractive hybridization. The cDNA from the cultures at the stage of somatic embryo maturation of embryogenic cell line Y35 of L. leptolepis L. princi-pis-rupprechtii was used as the tester and the cDNA from its subcultured callus was used as the driver. Eight hundreds ran-domly selected positive clones were sequenced, and 468 UniGenes were obtained finally.

为研究落叶松体细胞胚胎发生的分子机理，文章以日本落叶松华北落叶松杂种无性系胚性细胞系Y35体细胞胚成熟阶段培养物的cDNA为实验组，继代培养阶段胚性愈伤组织的cDNA为对照组，利用抑制性消减杂交技术(Suppression subtractive hybridization， SSH)构建了体细胞胚成熟阶段的差异表达基因文库。

To study the molecular mechanism of Larix somatic embryogenesis, a differentially expressed cDNA library of Larix somatic embryo in the period of maturation was constructed using suppression subtractive hybridization. The cDNA from the cultures at the stage of somatic embryo maturation of embryogenic cell line Y35 of L. leptolepis × L . princi-pis-rupprechtii was used as the tester and the cDNA from its subcultured callus was used as the driver. Eight hundreds ran-domly selected positive clones were sequenced, and 468 UniGenes were obtained finally.

为研究落叶松体细胞胚胎发生的分子机理，文章以日本落叶松×华北落叶松杂种无性系胚性细胞系Y35体细胞胚成熟阶段培养物的cDNA为实验组，继代培养阶段胚性愈伤组织的cDNA为对照组，利用抑制性消减杂交技术(Suppression subtractive hybridization， SSH)构建了体细胞胚成熟阶段的差异表达基因文库。

I am accused of being overreligious," she said in her quiet, frank manner,"but that does not prevent me thinking the children very cruel who obstinately commit such suicide.""

客人们在卡罗利娜·埃凯家里，举止就文雅一些，因为卡罗利娜的母亲治家很严厉。

Designed by French fashion house Herm è s, this elegant uniform was manufactured in our home, Hong Kong, and was the first without a hat.

由著名品牌 Herm è s 设计，这件高贵的制服是香港本土制造，是我们第一套不配帽子的制服。