次培养基

The plasmid of pEGFP-N1 was preserved in our laboratory.②The rat vibrissa follicles were dissected under a stereomicroscope. The dermal sheath was moved by incubated in dispase. The bulge regions of the hair follicles in anagen phase were carefully cut between the arrector pili muscle and the sebaceous gland, and then incubated with a mixture of trypsin and EDTA. The cell suspension was selected, and cultured in 10% fetal bovine serum DMEM/F12 FAD medium. After 7 days culture, HFSC was obtained by rapid adhering on collagen Ⅳ.

实验方法：大鼠在体视显微镜下分离出含真皮鞘的完整毛囊，dispase消化，将毛囊从真皮鞘中挤出，收集形态完好且处于生长期的毛囊，分别在毛球部上端、皮脂腺下端横切毛囊，取中间部分置于胰酶和EDTA中联合消化，向所得细胞悬液中添加含10%胎牛血清DMEM/F12完全FAD培养基，常规培养7 d后，采用IV型胶原快速贴壁法两次筛选以分离纯化大鼠毛囊干细胞。

Sub-culturing two bacteias in the same medium was futile, and third sub-culture slants showed no growth. attempts were made to separete these bacteria anaerobically with no success.

在相同培养基中接种两种细菌失败，第三次接种的斜面上无细菌生长，在无氧状态下试图分离这些细菌也以失败告终。

METHODS: Bone of limbs were obtained under sterile condition, washed by PBS, centrifugalization, and dropwise into centrifuge tube containing 1.073 g/mL Percoll separating medium, centrifugalization, and then adding L-DMEM containing 10% fetal bovine serum, benzylpenicillin sodium and phytomycin, cultured in a incubator at 37 ℃ with 5% saturated humidity. The culture medium was renewed after 48 hours, and non-adhesive cells were removed.

无菌条件下截取流产胎儿双侧四肢骨，PBS冲洗髓腔，离心去脂肪及上清液，L-DMEM重悬细胞，沿管壁缓慢滴加至底部有等量1.073 g/mL Percoll分离液的离心管中，离心后吸取中间界面白膜层，加入含体积分数为10%的胎牛血清、青霉素钠、链霉素的L-DMEM完全培养基，接种后置于37 ℃、体积分数为5%的CO2饱和湿度孵箱内培养。48 h后换液，去除未贴壁细胞，以后每2 d换液一次。

The puerperant and her relatives all signed informed consents. METHODS: Bone of limbs were obtained under sterile condition, washed by PBS, centrifugalization, and dropwise into centrifuge tube containing 1.073 g/mL Percoll separating medium, centrifugalization, and then adding L-DMEM containing 10% fetal bovine serum, benzylpenicillin sodium and phytomycin, cultured in a incubator at 37 ℃ with 5% saturated humidity.

无菌条件下截取流产胎儿双侧四肢骨，PBS冲洗髓腔，离心去脂肪及上清液，L-DMEM重悬细胞，沿管壁缓慢滴加至底部有等量1.073 g/mL Percoll分离液的离心管中，离心后吸取中间界面白膜层，加入含体积分数为10%的胎牛血清、青霉素钠、链霉素的L-DMEM完全培养基，接种后置于37 ℃、体积分数为5%的CO2饱和湿度孵箱内培养。48 h后换液，去除未贴壁细胞，以后每2 d换液一次。

Result Cells growed well in DMEM that contain 15% fetal cattle serum, and inoculative density was 4×10^4cellIml, penicillin and streptomycin concentration was 0.05%, pH was 7.2. It was easy to control and have good digest effect that use Hanks washes the cells three times advance then apply 0.05% trypsin-EDTA-Na2H2O to digest for one minute when convergence degree reaches 95-100%.

结果 含15%胎牛血清的DMEM培养基，接种密度为4×10^4个/mL，双抗浓度为0.5%，pH为7.2；在汇合度达95~100%时，Hanks洗3次，0.05%胰酶－EDTA-Na2H2O消化1min，传代效果最好，条件易于控制，且细胞能顺利生长。

During the intermissive fed-batch process, the feeding medium was added 10 times separately to make the total amount of C and N sources equal to those added in the batch process, and the yield of maduramicin was improved to 9645 μgmL^(-1), which is 15.7% higher than that obtained during the batch process. In the continuous fed-batch process, the maduramicin yield reaches 10328 μgmL^(-1), which is 6.0% higher than that obtained in the intermissive fed-batch process, and is 29.4% higher than that obtained in the batch process.

在间歇补料分批发酵工艺中，分10次补加培养基，使碳、氮源的总浓度与分批发酵相同，马杜霉素发酵单位提高到9645μgmL^(-1)，比分批发酵提高了15.7%；在连续补料分批发酵工艺中，马杜霉素发酵单位可达10328μgmL^(-1)，比间歇补料分批发酵提高了6.0%，比分批发酵提高了29.4%。

Thirty specimens samples were randomly divided into three groups blank control,media control and inoculated m edia group, 10 specimens were used in each group under the aerobic conditions,Sa polos glucose agar media with candida albicans inoculated .

需氧环境将白色念珠菌接种到沙保罗葡萄糖琼脂培养基后，将试件贴附其表面每周转换传递1次，共10周。0.5g/L戊二醛消灭细菌清洗，2周后，用Minolta CR-100 型色度计观察。

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。

The friable embryogenic callus with small grains and without proembryo formation was screened in the process of continuous subcultures for 4~6 times on the medium with low content of sucrose from embryogenic cultures induced from immature embryos in litchi (Litchi chinensis Soon.). The well-scattered embryogenic cell suspensions could be rapidly established from the friable embryogenic callus in 2 liquid initial MS media respectively supplemented with 2,4-D 2 mg/L or 2,4-D 2 mg/L, KT1 mg/L and AgNO3 5 mg/L shaking at 100~120 r/min within 10-14 days.

荔枝幼胚诱导的胚性培养物在低糖条件下连续继代4~6次左右，可筛选到颗粒细小、不含原胚的松散型胚性愈伤组织；以这种松散的胚性愈伤组织作为起始材料，在附加2，4-D 2mg/L或2，4-D 2mg/L、KT1 mg/L、AgNO3 5mg/L的MS液体启动培养基上振荡培养（100~120 r/min）10~14 d，即可建立起分散性良好的胚性悬浮细胞系。

Methods:Cardiac muscle of twoday neonate rattus was digested repeatedly by 0.0625% trypsin.The cardiocytes were collected in DMEM medium,which contains 10% fetal bovine serum.Myocardial cells were isolated with the technique of differential anchoring velocity with 90 min and were pured by Brdu.The cardiocytes were placed into CO2 incubator to incubate seven days after their purification.

应用0.0625%的胰蛋白酶重复消化出生第2 d乳鼠的心肌组织多次，收集的细胞用含10%胎牛血清的DMEM培养基中和，用差速贴壁分离法分离，在以溴脱氧尿嘧啶纯化心肌细胞后置CO2培养箱孵育7 d。

In the United States, chronic alcoholism and hepatitis C are the most common ones.

在美国，慢性酒精中毒，肝炎是最常见的。

If you have any questions, you can contact me anytime.

如果有任何问题，你可以随时联系我。