次培养

3H］-TdR incorporation rate was used as an index of cell proliferation. Additionally, we investigated whether the ASMC proliferative responses to SP were modulated by preincubation for 2 h with GR71251, a specific antagonist of NK-1 receptor, and neomycin, a phospholipase C inhibitor, and nitrendipine, a blockader of Ca2+ channel, respectively.

细胞增殖实验研究：将平滑肌细胞以1×103/孔接种在96孔板内，含10% FCS-DMEM培养24 h，吸去上清，以无血清DMEM洗涤2次，加入该培养基培养48 h，同步化之后以无血清培养基继续培养48 h或分别加入含SP(10-7～10-5 mol/L)和NK-1受体激动剂［Sar9，Met(O2)11］-SP(10-7～10-5 mol/L)的无血清DMEM培养相同时间。

The result indicated:① In accordance with culturing in the medium containing MS, SH, N6 macroelement, respectively, the effect of MS or SH macroelement in the primary culture is better than that of N6 macroelement, and the effect based on N6 macroelement is the best from 5th to 9th subculture.

结果表明：①在含MS、SH、N6的大量元素的培养基上的初代培养，MS或SH的效果好于N6，愈伤组织第5~9次继代培养中，N6大量元素培养基的继代培养效果最好。

The results showed that mucor protease was a kind of enzyme of strong induction. In addition to temperature,moisture and culture time,the ability of mucors producing profeinase were also closely related to culture methods,for example,the enzyme activity increased greatly when mucor was cultured with wheat bran and the leaven was turned over once as a lot of spores appeared.

结果表明，毛霉蛋白酶是诱导性很强的酶类，毛霉的产酶条件除与温度、培养时间、相对湿度等有关外，还与培养方式密切相关，以麦麸培养毛霉，当长有大量孢子时翻曲1次，可大幅度提高酶活力。

In this experiment, repetitiously intermittent bead-to-bead transfer of cell was chosen to simulate the close and continuous process. During the transfer of 1:4 expansion (the ratio of fresh and confluent microcarriers) for 4 times, it was found that cell density remained stable, while the HA titer showed a slight decrease probably due to the bad culture environments.

由于受实验条件的限制，本实验采用多次间歇加料的方式来模拟连续培养过程，经过4次新老载体1：4比例的放大培养，发现细胞密度可以保持稳定，而病毒产量有轻微下降的趋势，这可能是由于病毒生长环境比较恶劣的缘故。

Plantlet regeneration from cotyledon of Citrullus lanatus cv. Zhangkang No. 4 was studied. The results showed that aseptic seedlings should be cultured in dark for 3d, then exposed to a photoperiod of 16/8h/d for 3d. The callus induction rate was higher in cotyledons placed facing downwards to the medium than in those facing upwards. The highest induction rate occurred in MS medium containing 6-BA (2.0mg/L), kinetin (1.0mg/L)and GA3 (1.0mg/L). The induction initiated in dark and took 7 days, while callus growth and differentiation proceeding for 7 days under 16/8h light-dark cycles. A highest rate of embryogenic callus was obtained after 3 successive subcultures.

以无子西瓜郑抗4号无菌苗子叶为外植体，进行了体细胞胚发生及植株再生的研究，结果表明，无菌苗应先进行3d暗培养，然后采取光照16h/d和黑暗8h/d培养3d；将子叶的叶面朝下放置于培养基上的愈伤组织诱导率高于叶面朝上的培养方式；子叶诱导胚性愈伤组织的最适培养基配方为MS+6-BA2.0mg/L+KT1.0mg/L+GA31.0mg/L；诱导需要在黑暗条件下启动，进行7d暗培养，而生长分化于光照16h/d和黑暗8h/d条件下培养7d；继代3次得到的胚性愈伤组织最多；最适生根培养基配方为1/2MS+IBA0.3mg/L。

Factors that influence percentage of transformation including time of preculture, dipping and coculture, type and content of antibiotic, day of delayed selection culture, intension of selection were studied. The effective transformation system of "Starkrimson" was developed as follow: Leaves precultured in darkness for three days were dipped into agrobacterium suspension for three minutes, then were cocultured in darkness on MS medium with TDZ0.5mg/L and NAA 0.3mg/L for three days, and then cultured in darkness on MS medium with TDZ0.5mg/L NAA0.3mg/L and Cef 250mg/L.

通过对叶片预培养时间、侵染时间、共培养时间、抑菌素种类和浓度、延时筛选时间、不同选择压等遗传转化影响因素的研究，建立的'新红星'苹果遗传转化体系为：叶片黑暗预培养3天，经农杆菌侵染3分钟，在含TDZ0.5mg/L，NAA0.3mg/L的MS培养基上黑暗共培养3天，然后加250mg/L Cef黑暗维持培养3天，再进行Km12.5mg/L的选择压黑暗培养至抗性愈伤出现；转为光照条件1-2次的继代筛选，获得抗性芽后，进行扩繁。

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。

In the present study, we collected cumulus cells oocyte complex from ovaries of two different strain mice. The cumulusenclosed oocytes were cultured for 6 h in MEM supplemented with growth factor and FSH. The meiotic maturation of these oocytes has progressed to pro-metaphse Ⅰ stage and the condensed chromosomes are visible under DIC microscope, metaphase Ⅰ spindle even can be detected under Polscope. The metaphase Ⅰ spindles of oocytes were exchanged under such microscopes. After electric stimuli, 91. 6% and 91. 6% karyoplasts-cytoplasm pairs were fused respectively. The resulting oocytes were cultured further in MEM and over 80% of oocytes released the first polar body. 79% and 77% of oocytes formed two pronuclei after in vitro fertilization and the embryos were cultured in KSOM supplemented with amino acids. Over 60% of embryos developed to blastocyst stage.

在本研究中我们在取得两种不同品系小鼠的卵丘卵母细胞复合体后，先将卵丘卵母细胞复合体置于含有多种生长因子和激素的MEM培养液中培养6小时，此时卵母细胞已进入第一次减数分裂的前中期，并且在DIC倒置显微镜下可以看到浓缩的染色体，用Polscope可以发现明显的纺锤体，借助这种显微镜通过显微操作将两种不同品系小鼠来源的卵母细胞的MI纺锤体进行互换，经过三次直流电脉冲作用后，分别有91.6％的胞质—MI核质体对融合，经过进一步的培养后，超过80％的重组卵母细胞排出第一极体，体外受精后分别有79％和77％的重组卵形成双原核，受精后的胚胎在KSOM胚胎培养液中体外培养4天后，超过60％的胚胎发育至囊胚。

At the same time, group B cells were sequential planted by allowing cells to adhere to tissue culture dishes during two successive 2-hour incubations in order to remove non-adherent cells.

分别采用常规方法和序列黏附法培养。A组细胞于接种第4天洗去未黏附细胞，然后隔d换液1次；B组细胞在接种后每2h去除1次未黏附细胞，共2次。2组细胞均在第7天计数早期克隆，持续培养直到晚期克隆出现。

The results showed that after 2 h of culture in MEM the chromosomes of oocyte were seperated to form telophase I while a small spindle was observed around chromosomes of primary spermatocyte. However, two clear spindles were observed in the oocytes cultured in CB containing MEM. After further culture, the chromosomes of both primary spermatocyte and oocyte intermingled and formed one large spindle.

在无CB的培养液中培养的卵母细胞培养2小时后，卵母细胞已经进入第一次减数分裂的后期，染色体开始被拉向两极，而精母细胞的MI纺锤体才刚刚形成，虽然继续培养两者染色体可以合二为一并形成一个纺锤体，但是有些染色体发生滞后；当卵母细胞在含有CB的培养液中培养2小时后，在卵母细胞内形成两个相似大小的MI纺锤体，进一步培养形成一个大的纺锤体，染色体正常。

The absorption and distribution of chromium were studied in ryeusing nutrient culture technique and pot experiment.

采用不同浓度K2CrO4(0，0.4，0.8和1.2 mmol/L)的Hoagland营养液处理黑麦幼苗，测定铬在黑麦体内的亚细胞分布、铬化学形态及不同部位的积累。

By analyzing theory foundation of mathematical morphology in the digital image processing, researching morphology arithmetic of the binary Image, discussing two basic forms for the least structure element: dilation and erosion.

通过分析数学形态学在图像中的理论基础，研究二值图像的形态分析算法，探讨最小结构元素的两种基本形态：膨胀和腐蚀；分析了数学形态学复杂算法的基本原理，把数学形态学的部分并行处理理念引入到家实际应用中。