ATCC培养基介绍
安图生物普通培养基类(不含显色)产品简介
主要成分 基础营养 乳糖 指示剂 抑制剂 不分解乳糖
分解乳糖
产酸
PH 指示剂变红色
大肠杆菌等
红色大菌落 无色半透明小菌落
无明显变化 志贺菌等
痢疾杆菌
2.9
麦康凯琼脂平板
原理：该培养基利用胆盐来抑制革兰阳性细菌
生长，为中等程度选择性培养基，有少数革兰阴 性菌不生长；而对伤寒等沙门菌促进生长的作用。
2.12 物表测试平板
— 院内感染的检测卫兵
用途：物体表面细菌计数 培养环境中非苛养菌
底部计数方格设计 方便菌落计数
2.13 MH琼脂平板
用途：用于稀释法细菌药物敏感试验 （MIC和MBC测定）
原理：因M-H平板不含胸腺嘧啶，且 镁、钙金属离子含量低，不干扰磺胺 类药及氨基糖苷类药物的抑菌作用， 所以适宜细菌药物敏感试验的测定。
2.5 淋球菌琼脂平板
结果观察： 淋球菌ATCC19424生长良好（48h） 菌落较小，湿润，光滑，凸起，透明 无色成水珠状
2.6 巧克力色琼脂平板 （用于嗜血杆菌）
结果观察： 流感嗜血杆菌生长良好（48h）
2.7 沙保罗琼脂平板
用途： 用于培养霉菌和酵母菌等真菌 结果观察： 白色念珠菌生长良好；
原理：在45℃～50℃的基础培养基中加加入
血液（5～10%）可以保存血液中某些不耐热 的生长因子，同时血细胞不被破坏。若将NaCl 的浓度提高到0.85%,则可使血平皿经35℃18～24h后色泽 仍然鲜艳。在血平板上除可以观察菌落的形态外，还可以 判断溶血的情况。
结果观察：
化链，ATCC19615，ß溶血。 肺链，ATCC6303，ɑ溶血。 表皮，ATCC12228，不溶血。 金葡， ATCC29213，菌落黄色明显，ß溶血。
ATCC15834发根农杆菌使用说明
（必要时，可适当延长培养时间）。
菌 株 传 代 ：
将得到的菌株的新鲜培养物转接到适宜的固体培养基及液体培养基中（尽量
增大接种量：如用无菌吸管吸取≥50μl 新鲜培养物至固体培养基，边移动边缓
慢释放），适宜温度下培养，用以菌株的保藏、传代及制备工作菌株。
面蘸取少量涂板或进行液体培养即可。也可以完全融解后使用，但随着冻融次数
的增加，细菌的活力会逐渐下降。
2，为保证菌种纯正，避免其它细菌污染，尽量先划平板，然后再挑单克隆
菌落进行后续操作。
冷 冻 管 开 封 ：
用浸过 75%酒精的脱脂棉严格消毒冷冻管盖。
ATCC15834 发根农杆菌
编号
名称
北京华越洋生物 NRR01170 ATCC15834 发根农杆菌
基 本 信 息 ：
名称：ATCC15834 发根农杆菌
规格：300ul 甘油菌
储 存 温 度 ： -‐80℃
简 介 ：
注 意 事 项 ：
1、菌种活化前，将冷冻管保存在低温、清洁、干燥的环境中，长时间室温下放
置会导致菌种衰退；
2、冷冻管开封、冻干粉复溶、菌株恢复培养等操作应在无菌条件下进行；
3、一些菌种经过冷冻干燥保存后，延迟期较长，部分需连续两次继代培养才能
正常生长；
4、苛养菌的培养需采用含特定营养成分的培养基，敬请正确选择，不清楚时来
菌 株 复 溶 ：
无菌环境中旋开装有复溶液的滴瓶盖，吸取 1ml 左右复溶液，加入到冷冻
管中。轻轻振荡，使冻干菌株溶解呈悬浮状。
菌株复壮：
用无菌吸管吸取菌悬液，转移到复溶液滴瓶中。做好标识，在适宜温度下培
养。细菌在 30-‐35℃培养箱中培养 24-‐48h，真菌在 23-
金黄色葡萄球菌ATCC6538使用说明
金黄色葡萄球菌ATCC6538编号 名称北京华越洋生物NRR01280 金黄色葡萄球菌ATCC6538 基本信息：名称：金黄色葡萄球菌ATCC6538规格：300ul甘油菌类别： 金黄色葡萄球菌抗性： 无培养基： LB培养方式： 37℃，有氧保存方式： 30%甘油，-‐80℃菌株简介:在LB平板上菌落呈金黄色或为白色,大而突起，圆形，不透明，表面光滑，周围有溶血圈。
在Baird-‐Parker平板上呈现圆形，光滑凸起，湿润。
显色呈灰色到黑色，边缘为淡色。
经革兰氏染色显色其为革兰氏阳性。
经无菌水10倍稀释，涂平板，37℃培养过夜未见其他微生物生长，经可见金黄色葡萄球菌菌落。
操作方法：1. 从泡沫盒中取出菌冻干管，置于超净台中。
2. 去除冻干管上的封口膜，然后将冻干管在酒精灯上烧一下灭菌。
3. 准备好无抗性的LB平板，往冻干管中加入适量的LB培养基，用接种环四区划线，37℃过夜培养。
4. 挑取单菌落进行扩大培养。
5. 冻存菌株，做好标记。
冷冻管开封：用浸过75%酒精的脱脂棉严格消毒冷冻管盖。
菌株复溶：无菌环境中旋开装有复溶液的滴瓶盖，吸取1ml 左右复溶液，加入到冷冻管中。
轻轻振荡，使冻干菌株溶解呈悬浮状。
菌株复壮：用无菌吸管吸取菌悬液，转移到复溶液滴瓶中。
做好标识，在适宜温度下培养。
细菌在30-‐35℃培养箱中培养24-‐48h，真菌在23-‐28℃培养箱中培养24-‐72h （必要时，可适当延长培养时间）。
菌株传代：将得到的菌株的新鲜培养物转接到适宜的固体培养基及液体培养基中（尽量增大接种量：如用无菌吸管吸取≥50μl 新鲜培养物至固体培养基，边移动边缓慢释放），适宜温度下培养，用以菌株的保藏、传代及制备工作菌株。
注意事项：1、菌种活化前，将冷冻管保存在低温、清洁、干燥的环境中，长时间室温下放置会导致菌种衰退；2、冷冻管开封、冻干粉复溶、菌株恢复培养等操作应在无菌条件下进行；3、一些菌种经过冷冻干燥保存后，延迟期较长，部分需连续两次继代培养才能正常生长；4、苛养菌的培养需采用含特定营养成分的培养基，敬请正确选择，不清楚时来电询问；5、某些厌氧菌的培养，自开封到接种完成，均需以无氧气体充填，以保持厌氧状态；培养过程中亦要保持厌氧状态；6、某些菌种，如肺炎链球菌、流感嗜血杆菌、淋病奈瑟菌等需要5-‐10%CO2 促进生长；7、如发现冷冻管盖松动、复溶液浑浊等异常情况，应停止使用对应产品。
ATCC菌株操作说明中文版
TIB. 081
养基。在最初的 48 小时培养期内不要揭开接过种的平皿。
Candida sp. Capnocytophaga sp. Cellulosimicrobium sp. Chryseobacterium sp. Citrobacter sp. Cladosporium sp. Clostridium sp.
方法 8 缓冲活性炭酵母提取物琼脂-----35℃，正常环境，3-5
天。
方法 9 V 琼脂或巧克力琼脂-----35℃，5％－10％ CO2 环
境，48 小时。 方法 10 脑心浸出液肉汤，胰蛋白胨大豆肉汤，或 0.85％生
理盐水均可选用。 非选择性羊血琼脂或胰蛋白胨大豆琼脂-----35℃，正
中保存的。
Bacteroides sp.
拟杆菌属
方法 4
注： Bacteroides ureolyticus(解尿素拟杆菌)应培养 5 天，菌落很小。有时
需要多传 1 代以得到足够的供试菌落。
Bifidobacterium sp. Bordetella sp.
双岐杆菌属 博德特氏菌属
方法 4 方法 3
Aeromonas salmonicida(杀鲑气单胞菌 )应在 25℃培养。
Alcaligenes sp.
产碱杆菌
方法 1
Alicyclobacillus sp.
脂环酸芽孢杆菌 方法 12
Alloiococcus sp.
Method 2
Aneurinibacillus sp.
硫 胺 素 芽 孢 杆 菌 方法 1 属
方法 14 取 一 颗 冻 干 颗 粒 于 SP4 葡 萄 糖 肉 汤 中 ， 若 是
KWIKSTIK 则溶于自带的培养基中。作系列梯度稀 释（例如，1:2，1:4，1:8，1:16，1:32）。在 35℃下， 正 常 环 境中 培 养。 一 旦肉 汤 从红 色 变成 黄 色（ 1-4 周），转接 0.2 mL 到 SP4 葡萄糖琼脂，划线分离。 不要使用棉质拭子或木质棒。35℃，CO2 环境中， 可以使用烛缸，培养 5-15 天。使用显微镜观察平板 菌落。 方法 15
生物技术工业用培养基介绍知识资料
10个/包
80
用于环境、器皿、设备和表面的无菌检测
含青霉素酶接触皿培养基平板（6.5cm）
10个/包
80
用于环境、器皿、设备和表面的无菌检测
RODAC接触皿（TSA）（6.5cm）
10个/包
50
用于环境、器皿、设备和表面的无菌检测
NA平板（加青霉素酶）（9cm）
10个/包
灭菌。
（3）可以长时间培养。
（4）有效期3个月。
（5）根据客户要求定制灌装量，最大可以多达35mL。
（6）每块平板均标注名称/批号、有效期。
2、55m物表测试培养基
（1）储存温度：4-8℃。
（2）三层纸塑袋+真空袋，有效降低冷凝水产生，透气性好，阻菌性能好，辐射
灭菌。
（3）可以长时间培养。
（4）有效期3个月。
改良Skirrow氏琼脂平板（9cm）
10个/包
90
用于弯曲杆菌的分离培养
二、显色培养基系列
产品名称
百博色培养基平板
10块/包
110
用于沙门氏菌选择分离培养
李斯特氏菌显色培养基平板
10块/包
160
用于单增李斯特氏菌的选择分离培养
O157显色培养基平板
10块/包
130
用于大肠埃希氏O157菌的选择分离培养
MRS琼脂平板（9cm）
10个/包
120
用于食品中乳酸菌总数测定
质量保障
（1）进口原料，全自动生产线生产。
（2）一重保障：高压蒸汽灭菌。
（3）二重保障：万级洁净环境全封闭自动灌装、百级环境灌装。
（4）三重保障：钴60 辐射灭菌。
atcc标准细胞培养方法
atcc标准细胞培养方法
ATCC细胞培养方法是细胞培养的一种标准方法。
该方法包括以下步骤：
1. 选择适当的培养基：根据细胞种类选择适当的培养基，如DMEM、RPMI、MEM等。
加入10%的胎牛血清以及必要的抗生素和抗真菌药物。
2. 准备细胞：从冻存细胞库中取出细胞，放置在无菌条件下，用PBS（含有钙和镁）洗涤细胞2-3次，去除冻存液。
3. 接种细胞：将细胞转移到含有培养基的无菌培养皿中。
培养皿容积应根据细胞数量和培养时间来确定。
4. 细胞培养：将培养皿放置在37℃的培养箱中，保持湿润的环境。
定期更换培养基，一般每两天更换一次。
注意细胞密度的控制，不要过度密集。
5. 细胞分离：在培养达到一定密度后，将细胞用胰酶或EDTA等酶类分离剂进行消化，分离到新的培养皿中。
6. 冻存细胞：当细胞密度达到适当的水平后，使用液氮冻存细胞，以备以后使用。
注意事项：培养过程中必须严格控制消毒，确保细胞无污染；定期检查细胞状态、生长情况、污染情况等，及时处理异常情况。
atcc标准菌株说明书
atcc标准菌株说明书
1、ATCC菌种批号说明:菌名称简称的拼音首字母+转种日期-代数-编号。
2、菌株每转种一次即增加一代，实验用应控制在五代以内。
3、购买菌株后，立即进行转种，不宜长期冷藏保存。
4、转种方式:
普通菌斜面:用接种环挑取斜面上的菌落，转种至适宜的液体培养基中，按药典规定的时间培养后，将液体培养物分装至无菌冻存管中，每支 1.5~2.0ml，冰箱冷冻(-20°℃)保存。
每次实验取一支转种后使用。
如:生孢梭菌:将小管中液体培养物接入硫乙醇酸盐液体培养基中，ATCC菌种按药典规定的时间培养后，将液体培养物分装至无菌冻存管中，每支1.5~2.0ml，冰箱冷冻(-20℃)保存。
每次实验取一支转种后使用。
黑曲霉:用接种环蘸取小管中孢子悬液，并涂布至改良马丁琼脂斜面上，培养1周后，按药典方法将孢子洗下，孢子悬液冰箱冷藏(4℃)保存。
实验时可直接取该悬液稀释至合适的级别后使用。
5、转种的液体培养基体积推荐为100ml，可分装约40~50支无菌冻存管，平时使用时应定期进行菌株的鉴定，若菌落形态不典型或特征反应不明显，说明该ATCC菌种已不适宜继续使用，应销毁后购买新菌株。
BEAS-2B所用培养基
一、BEBM培养基（ATCC）(1个)1.African Dust Storms Reaching Puerto Rican Coast Stimulate the Secretion of IL-6 and IL-8 and Cause Cytotoxicity to Human Bronchial Epithelial Cells (BEAS-2B)BEAS-2B cells were obtained from the American Type Culture Collection (ATCC, Manassas, V A, USA; Cat No CRL-9609). Cells were cultured according to ATCC protocols,maintained in keratinocyte basal medium (KBM-2, Lonza, Walkersville, MD, USA Cat No CC 3103) and supplemented with keratinocyte growth medium (KGM-2 SingleQuots; Lonza, Walkersville, MD, USA; Cat No CC4152). Cells were used at passages 44 - 59 and maintained at 37°C in a humidified atmosphere of 5% CO2二、DMEM（11）1.Diatom-Derived Polyunsaturated Aldehydes Activate Cell Death in Human Cancer Cell Lines but Not Normal CellsIn an independent experiment, A549 cells (2×103cells well-1) were seeded in a 96-well plate and kept overnight for attachment. The next day the medium was replaced with fresh medium with three concentrations (2, 5 and 10 µM) for each of three PUAs (DD, OD, and HD, Sigma-Aldrich Inc., Milano, Italy) tested and with caspase-3 Inhibitor (C30H41FN4O12, sc-3075, Santa Cruz) at 9.7 µM; cells were allowed to grow for 24, 48 and 72 h. After aldehyde treatment, viable cells were evaluated as described below.The BEAS-2B (ATCC CRL-9609) lung/brunch normal epithelial cell line was maintained in DMEM (Dulbecco's modified Eagle's medium) supplemented with 50% fetal bovine serum (FBS), 100 units ml−1 penicillin and 100 µg ml−1streptomycin. Cells were incubated in a 5% CO2humidified chamber at 37°C for growth. BEAS-2B (2×103 cells well−1) was seeded in a 96-well plate and kept overnight for attachment. The next day the medium was replaced with fresh medium with three concentrations (2, 5 and 10 µM) for each of three PUAs (DD, OD, and HD, Sigma-Aldrich Inc., Milano, Italy) tested; cells were allowed to grow for 24, 48 and 72 h. After incubation, the supernatant was removed and adherent cells were examined for viability.2.Gene Expression Profile and Toxic Effects in Human Bronchial Epithelial Cells Exposed to ZearalenoneHuman bronchial epithelial BEAS-2B cell line [19] (from the American Type Culture Collection, ATCC) was cultured in Dulbecco's Modified Eagles Medium(DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS) and 100 U/ml of penicillin and 10 µg/ml of streptomycin. All cells were maintained in a 37°C humidified incubator with 5% CO2. DMEM with Geneticin (G418, 200 µg/ml) were used to maintain and select Cygb overexpressing cells.3.Arsenic-induced sub-lethal stress reprograms human bronchialepithelial cells to CD61¯ cancer stem cellsThe human bronchial epithelial cell line BEAS-2B was purchased from the American Type Culture Collection (ATCC, Manassas, V A) and cultured in DMEM with 10% FBS. For As3+treatment, BEAS-2B cells were treated with 0.25 μM NaAsO2 continuously for six months. Cells cultured without As3+treatment for six months were used as parental cell control in all experiments. Cell transformation was evaluated by using in vitro soft agar colony-forming assay and in vivo tumorigenicity experiment in athymic nude mice. For the soft agar colony-forming assay, 1% agar was melted and mixed with 2 ×DMEM m edium in a 1:1 ratio to produce a supporting layer (0.5%) in a 6-well tissue culture plate (1 ml/well). The bottom agar layer was allowed to solidify at room temperature for 20 min. The top layer containing 0.3% agar (1 ml/well) was prepared by mixing stock agar solutions with 5,000 cells in 2× DMEM medium and was laid on the top of the bottom agar layer. Fresh DMEM medium was added every 3 days. Three weeks later, colonies were stained with a 0.05% crystal violet solution, counted, and photographed under a microscope4.Receptor Interacting Protein-2 Plays a Critical Role in Human Lung Epithelial Cells Survival in Response to Fas-Induced Cell-DeathHuman lung epithelial cells (BEAS-2B) were purchased from ATCC (ATCC CRL9609). BEAS-2B cells were cultured in DMEM (MediaTech, Inc) supplemented with 10% heat-inactivated FBS (Atlanta Biologicals) and 1% penicillin-streptomycin (Invitrogen Life Technologies). Anti-Fas (human, activating), clone CH11 was purchased from Upstate Cell Signaling Solution, USA. Cells were regularly checked to confirm the absence of Mycoplasma contamination [16]. Cell culture medium was used for detection of LDH release as a cell death signature. Cell lysates were analyzed for proteins by immunoblots.5.NEIL2 Protects against Oxidative DNA Damage Induced by Sidestream Smoke in Human CellsHuman BEAS-2B lung epithelial cells were from the American Type Culture Collection (ATCC, Manassas, V A). Human pulmonary fibroblasts (hPF, ScienCell, Carlsbad, CA) and human embryonic kidney 293 (HEK293) cells were grown at ambient oxygen levels and 10% CO2i n DMEM supplemented with 10% fetal calf serum before treatment. Our laboratories have been using HEK293 cells for many transfection-based experiments, therefore, this cell line was used along with the two pulmonary cell lines. Cells were treated with various concentrations of SSS for 24 h before collecting by trypsinization.6.MicroRNA-34c is associated with emphysema severity and modulates SERPINE1 expressionBEAS-2B and HFL1 were cultured in RPMI and DMEM,respectively, supplemented with antibiotics and 10% FCS and incubated in 5% CO2.7.Chronic Arsenic Exposure and Angiogenesis in Human Bronchial Epithelial Cells via the ROS/miR-199a-5p/HIF-1α/COX-2 PathwayCell culture and generation of stable cell lines. Human bronchial epithelial BEAS-2B cells [American Type Culture Collection (ATCC), Manassas, V A, USA] were cultured in DMEM (Dulbecco’s modified Eagle’s medium; Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS).8. Differential responses to genotoxic agents between induced pluripotent stem cells and tumor cell linesMitomycin C treated murine embryonic fibroblasts (MEFs) were prepared as feeder cells. Tera-1 cells obtained from American Type Culture Collection (A TCC) were cultured in McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS). BEAS-2B cells obtained from ATCC were cultured in DMEM supplemented with 10% FBS.9The effect of particle size, location and season on the toxicity of urban and rural particulate mattert he bronchial epithelial cell line (BEAS-2B) was obtained from the American Type Culture Collection (ATCC, Rockville, MD) and maintained in DMEM medium (Dulbecco’s Modified Eagle Medium; Gibco) with 10% FBS (Gemini Bio Produ10.Perilla frutescens Leaf Extract Inhibits Mite Major Allergen Der p 2-induced Gene Expression of Pro-Allergic and Pro-Inflammatory Cytokines in Human Bronchial Epithelial Cell BEAS-2BThe nontumorigenic human bronchial epithelial cell BEAS-2B (ATCC®CRL-2503™) was obtained from American Type Culture Collection (ATCC; Rockville, MD, USA) and cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% v/v fetal bovine serum (Gibco BRL, Gaithersburg, MD, USA) and 100 µg/mL penicillin/streptomycin (Sigma, St. Louis, MO) at 37°C ina humidified atmosphere containing 5% CO2.11.Sodium metavanadate exhibits carcinogenic tendencies in vitroin immortalized human bronchial epithelial cells†Immortalized human bronchial epithelial cells (Beas-2B; #CRL-9609, ATCC, Manassas, V A) were cultured in 1×Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals, Lawrenceville, GA) and 100 ug mL−1 Pen Strep (GIBCO, Grand Island, NY) (“complete culture media”). The cells weremaintained in 25 cm2polystyrene tissue culture flasks in an incubator at 37°C with 5% CO2 and 100% humidity. The media were changed every 2 days, and the cells were passaged every 4 days using 0.05% trypsin-EDTA (GIBCO).三、F12 （1）1.Biological responses according to the shape and size of carbon nanotubes in BEAS-2B and MESO-1 cellsBEAS-2B cells were cultured in Ham’s Nutrient Mixture F-12(Nacalai Tesque) with 10% fetal bovine serum ([FBS] Life Technologies, Carlsbad, CA, USA), and MESO-1 cells were cultured in RPMI1640 supplemented with 10% FBS.四、DMEM·F12 (3)1.Effect of Tumor Necrosis Factor Family Member LIGHT (TNFSF14) on the Activation of Basophils and Eosinophils Interacting with Bronchial Epithelial CellsBEAS-2B cells were grown in Dulbecco's modified Eagle's medium nutrient mixture F12 (Invitrogen) with 10% FBS at 37°C in a humidified 5% CO2 atmosphere until confluence to cell monolayer. In coculture, the medium of BEAS-2B cells was replaced with RPMI-1640 medium containing 10% FBS (Invitrogen) with or without basophils/eosinophils. For inhibition experiments, basophils/eosinophils and BEAS-2B cells were pretreated with signaling molecule inhibitors for 1h before coculture and treatment by LIGHT.2.Sphingosine 1-Phosphate (S1P) Induced Interleukin-8 (IL-8) Release Is Mediated by S1P Receptor 2 and Nuclear Factor κB in BEAS-2B Cells Human BEAS-2B cells (ATCC, Manassas, V A) were grown in DMEM:F12 10%FBS 100 U/ml penicillin, 100 µg/ml streptomycin and 2500 ng/ml amphotericin B (PSA) (Invitrogen, Carlsbad, CA, USA) in 75 cm2tissue culture flasks at 37°C and 5% CO2. Culture medium was changed every 2 days and cells were seeded into new flasks when approximately 80% confluent. Cells were detached by incubation with 0.25% trypsin (Sigma-Aldrich). For experimentation, cells were seeded in 6 well plates at a density of 50 000 cells per well and grown for 3 days in culture medium. Cells were serum-starved for 24 hours in DMEM:F12 0.1%BSA (Sigma-Aldrich) with PSA. Starvation medium was changed prior to all experiments and culture supernatant was collected at the end of the incubation period.3.EFFECTS OF COREXIT DISPERSANTS ON CYTOTOXICITY PARAMETERS IN A CULTURED HUMAN BRONCHIAL AIRWAY CELLS, BEAS-2BCommercially available COREXIT 9500A, 9527A, and 9580A dispersants asliquid solutions were provided by a contract between Nalco/Exxon Energy Chemicals, L.P. (Sugar Land, TX) and Tulane University (New Orleans, LA). Fibronectin, collagen, Dulbecco’s modified Eagle’s medium with nutrient mixture (DMEM/F-12),and penicillin/streptomycin were purchased from Sigma-Aldrich (St. Louis, MO).五、LHC-9（中科院）(6)1.Transforming growth factor-β impairs glucocorticoid activity in theA549 lung adenocarcinoma cell lineBEAS-2B cells (from ATCC, Manassas, V A, USA) were cultured in LHC-9 media containing 2% vv−1heat-inactivated FCS, 15 mM HEPES, 0.2% vv−1sodium bicarbonate, 2 mM L-glutamine, 50 IU·mL−1penicillin and 50 µg·mL−1 streptomycin and maintained at 37°C in a humidified 5% CO2/95% air atmosphere. For experiments, the cells were seeded at 25 000 cells/well in 24 well plates and incubated in serum-free medium (phenol red-free Roswell Park Memorial Institute (RPMI) containing 15 mM HEPES, 0.2% vv−1sodium bicarbonate, 50 IU·mL−1 penicillin, 50 µg·mL−1streptomycin, 0.25% wv−1 BSA, 2 mM L-glutamine, 1% v/v sodium pyruvate and 1% v/v non-essential amino acids) for 24 h prior to treatment with TGF-β(40 pM) for 24 h. The cells were then treated with dexamethasone. IL-1α(1 ng·mL−1) was added 30 min after the addition of dexamethasone to stimulate the production of cytokines when required. Cells were supplemented with Monomed A (1% vv−1) 24 h after culture in serum-free medium.2.Bronchial epithelial cells are rendered insensitive to glucocorticoid transactivation by transforming growth factor-β1A549 lung adenocarcinoma and BEAS-2B bronchial epithelial cell lines were cultured as described [9]【[9]就是上面的文章】3.Low-solubility particles and a Trojan-horse type mechanism of toxicity: the case of cobalt oxide on human lung cellsThe cytotoxicity of cobalt oxide particles was evaluated on BEAS-2B cells by measuring the CellTiter-Glo assay after 72 h exposure to increasing concentrations of Co3O4particles in LHC9 (0–1000 μg.mL−1Co). Because of the low level of toxicity of cobalt oxide particles [26], we investigated a wide range of concentrations. In parallel, the cytotoxicity of LB-3 latex beads was evaluated for similar concentrations (0–1000 μg.mL−1). After 72 h exposure, the sedimentation of 400 nm cobalt oxide particles, as well as latex beads, was fully achieved which validate the cytotoxicity results obtained for these particles suspensions, all the particles being available to the adhering cells.4.Curcumin and Vitamin E Protect against Adverse Effects ofBenzo[a]pyrene in Lung Epithelial CellsHuman bronchial epithelial cell line, BEAS-2B, was obtained from American Type Culture Collection (ATCC). The cells were cultured as described in our previous study [31]. In brief, after the flasks/dishes/plates were coated, BEAS-2B cells were cultured in LHC-9 (Life Technology, MI) completed growth medium containing 50U penicillin/mL and 50 μg/mL streptomycin. The cells were then incubated at 37°C in 95% air and 5% CO2. At 80% confluence, cells were harvested using 0.05% trypsin/EDTA solution (HyClone) and were then sub-cultured.5..6.. Regulation of CYP3A genes by glucocorticoids in human lung cells BEAS-2B cells (American Type Culture Collection) were cultured i n LHC-9 medium (Life Technologies). Lobar cells (donor number 01334) were cultured in BronchiaLife Basal Medium supplemented with the BronchiaLife B/T supplement kit (Lifeline Cell Technology, Walkersville, MD).六、BEGM (2)1.2.Brd4 Is Essential for IL-1β-Induced Inflammation in Human Airway Epithelial CellsNormal human bronchial epithelial cells (NHBE) of non-smoking subjects were obtained from Lonza (Lonza, Slough, UK) and grown in bronchial epithelial cells growth medium (BEGM) supplemented with growth supplements (SingleQouts kit, Lonza) as recommended by the manufacturer. Cells were passaged at passages 2–8 using the ReagentPack subculture kit (Lonza) following suppliers instructions. Cells were cultured until 80% confluent at 37°C and 5% CO2. Prior to experiments, monolayers of cells (70–80% confluence) were incubated in basal medium (supplement free) overnight. Cells were treated with BET inhibitors (JQ1 and PFI-1) prior to stimulation with IL-1β (1 ng/ml) in the presence or absence of H2O2 (100µM). All experiments were performed at least four times.七．1640 (2)1.Respiratory Syncytial Virus Infection, TLR3 Ligands, and Proinflammatory Cytokines Induce CD161 Ligand LLT1 Expression on the Respiratory EpitheliumThe transformed human bronchial epithelial cell line BEAS-2B (ATCC and LGC Standards, United Kingdom) was cultured in RPMI 1640 m edium supplemented with penicillin (100 U/ml), streptomycin (100 μg/ml), L-glutamine (0.8 mM) (culture medium), and 10% heat-inactivated fetal calf serum (FCS) (all from PAA Laboratories).2.Prolonged cigarette smoke exposure alters mitochondrial structure and function in airway epithelial cellsBEAS-2B cells were grown in RPMI 1640(Lonza, Walkersville, MD) supplemented with 15% heat-inactivated Fetal Bovine Serum (FBS), penicillin (100 U/ml) (Lonza) and streptomycin (100 μg/ml) (Lonza) on collagen-coated plates. Cells were grown to >90% confluence and passaged by trypsin.。
ATCC简介
ATCC简介
美国菌种保藏中心，又称美国模式菌种收集中心(ATCC)，是位于马里兰洲洛克菲勒的一家私营的，非赢利性组织。
目前它可以提供各种动植物细胞、标准品、菌株达两万多种。
ATCC的成立可追溯到1925年，一个科学委员会认识到科学研究对于微生物的大量需求而萌发了创建一个微生物相关制品供应中心的想法。
在生物制品方面，我公司是世界卫生组织下属生物标准品实验室（英国生物制品检定所NIBSC和荷兰VQC）、欧洲药品质量监察会（EDQM）在中国指定的进口商，同时我部门也在代理进口一些其他菌种保藏机构的产品，比如德国国家菌种保藏中心(DSMZ)、英国国立标准菌种收藏中心(NCTC)等，为国内科研以及生产用户提供细胞株、菌株等生物标准品。
ATCC 美国模式培养物集存库简介
ATCC成立于1925年，是世界上最大的生物资源中心，由美国14家生化、医学类行业协会组成的理事会负责管理，是一家全球性、非盈利生物标准品资源中心。
ATCC向全球发布其获取、鉴定、保存及开发的生物标准品，推动科学研究的验证、应用及进步。
目前，ATCC有29,000多种不同品系可靠的动物细胞和微生物培养体，它能满足各科学团体对可信赖品系
的需要。
ATCC是一个非官方(学会单位)无利润的公司，由各科学组织推选成员组成理事会进行管理。
现
有工作人员125人。
在ATCC可利用各种材料以及工具和设备进行广泛的研究。
抑菌实验方案
抑菌实验方案一、菌种大肠杆菌ATCC25922 金黄色葡萄球菌ATCC6538二、培养基LB培养基：胰蛋白胨10g/L、酵母膏5g/L、氯化钠10g/L、琼脂15~20g，用10mol/LNaOH调pH，范围7.0~7.4倒平板（平板培养基）1.将培养皿洗净、干燥，使各培养皿盖方向一致，用牛皮纸包好，或放入特制铁罐内，注明皿盖的方向。
高压灭菌或干燥灭菌后备用。
2.取刚灭菌后尚未凝固的培养基置于无菌箱或超净工作台内，先左手持三角瓶,右手反转手掌用中指和无名指拨出棉塞或硅胶赛(或不翻转手掌用小指和手掌拨出棉塞) ,同时将三角瓶转换至右手，而后左手拿平皿,以大拇指和中指将皿盖打开一缝，至瓶口刚好伸入。
三角瓶口经火焰烧灼后，倾入灭菌或溶化、冷却至55~60°C的培养基约15mL(勿使瓶口靠在平皿壁上，以免沾染皿壁)，迅速盖好皿盖，置于桌上轻轻旋转平皿,使培养基均匀分布于整个平皿底部,冷凝后即为平板培养基。
有时凝固后的培养基表面有冷凝水，可将平皿盖打开倒置于37℃温箱，除去冷凝水，否则将影响平板分离培养效果。
为防止冷凝水过多，也可将培养基冷却至45 ~ 50°C再倒平板。
培养基在30℃下放置一天，无污染的即可使用。
一般用牛皮纸包裹好存放于2-8℃冰箱中备用三、器械培养皿（90mm）、滤纸、三角耙、接种环、试管四、实验步骤（一）、抑菌圈的测定(滤纸片法)1、菌种接种将大肠杆菌、金黄色葡萄球菌反复转种2次，使菌种新鲜，置30℃恒温培养箱培养1d，备用。
2、菌悬液制备在超净台里将转接后生长良好的菌种试管斜面注入适量无菌水(约5mL)，接种环轻刮菌落，摇晃混匀。
3、涂布在超净台里用枪取0.1ml菌悬液注入平板，放置5min左右后用三角耙涂布均匀，每种菌设置两个重复平板4、贴片在超净台里将平板平均分为6个区，对角设置平行组，同时用无菌水做对照组,硫酸链霉素(10U)作阳性对照。
每两张滤纸片(直径1.1cm或0.6cm)为一贴，用镊子夹着滤纸片在样品中轻轻蘸一下后，取出贴在平板的指定位置，轻摁一下使滤纸片牢固，置(正放)4℃冰箱中放24h。
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ATCC high-performance media and sera are uniquely formulated according to cell-growth recommendations of original cell line depositors and ATCC cell culture specialists.Using ATCC high-performance media and sera ensures robust cell growth with minimum cell loss especially when reviving cells from cryopreservation. With ATCC media and sera cell cultures grow in a continuous and consistent manner providing a ready supply of cells when needed.Your Discoveries Begin With Us.®TMATCC ® High-PerformanceMedia and SeraOur lab has used media and sera from many companies and has found ATCC formulations to be consistently top caliber. Cryopreserved cells recover much faster and viability is higher saving our lab both time and money.—Shannon Reagan-Shaw, Research Specialist, University of Wisconsin-Madisonrequirements for individual cell lines for optimal cell culturing.ATCC scientists subject cell lines to comprehen-sive evaluations as part of the cell line accession-ing process. During these tests, cell lines are fully authenticated and growth parameters are defined to promote uniform and stable growth over severalsera requirements, which are tested and used on ATCC cell lines.The ATCC Cell Culture Facility uses ATCC high- performance sera and media to produce and distribute over 3,600 different cell lines. These reagents are made available to the research com-munity only through ATCC.We have cultured a large number of different cell types and have used many different ATCC media formula-tions with 100% success. For us, ATCC media has taken the worry out of cell culture.—Lisa Ryner, Ph.D., Research Scientist, KAI PharmaceuticalsATCC high-performance animal sera come from USDA-approved origins. Experienced technicians thoroughly test the packaged sera to eliminate contamination concerns. All ATCC sera are prequali-ATCC High-Performance SeraProduct name Catalog No. Volume Fetal Bovine Serum 30-2020 500 ml30-2021 100 ml Fetal Bovine Serum, ES Qualified* SCRR-30-2020 500 ml Calf Bovine Serum 30-2030 500 ml30-2031 100 ml Horse Serum 30-2040 500 ml30-2041100 mlATCC sera result in the most reliable cell growth — experience the differencefied on multiple cell lines to ensure robust growth using both sequential subcultures and plating efficiencies.• Each lot of ATCC high-performance sera is subjected to comprehensive tests for sterility and performance.• Sterility tests are performed on each lot using current USP methods for bacterial and fungal contamination.• Comprehensive viral testing adheres to 9 CFR 113.53 guidelines.• Mycoplasma testing is performed via direct culture and Hoechst DNA staining.• Limulus amoebocyte lysate (LAL) procedure is used to measure endotoxin values.• All tests are performed post-packaging to guarantee the highest-quality, best- performing sera.* Qualified for mouse and human embryonic stem cells by the ATCC Stem Cell CenterAnimal Sera product listare based on extensive evaluation of growth and viability requirements of individual cell lines.All ATCC products are manufactured to exact specifications and each lot is rigorously tested to meet the standards of performance and quality imposed by the ATCC Cell Culture Facility, where distribution stocks for numerous cell lines are pro-duced routinely.disturb normal cell growth.The table on the following page describes the dif-ferences that separate ATCC high-performance me-dia from generic formulas.After a couple of failed attempts to grow ZF-4 cells in medium supplemented with serum from other manufacturers, I switched to ATCC Fetal Bovine Serum (Catalog no. 30-2020). Using this serum, I obtained good recovery, viability and robust growth.—Dr. Annette Schenck, Max Planck Institute of MolecularCell Biology and Genetics, Dresden, GermanyProduct name Catalog No. Key Components Uses and BenefitsRPMI-1640 Medium30-2001(500 ml)1 mM sodium pyruvate2 mM L-glutamine4500 mg/l glucose10 mM HEPES1500 mg/l sodium bicarbonateBalanced energy sources to serve as carbonskeletons for anabolic processes as well as forprotein production and nucleic acid metabolismwhile limiting toxic ammonia build-up.Optimized to maintain pHTo be used with 5% CO2to maintain pHATCC Media product list* * See ATCC website for complete media formulations.ATCC Cell Culture ReagentsCustomize growth conditions with ATCC-tested media supplements and antibioticsCell culture reagents product listProduct name Catalog No. Uses and BenefitsL-Glutamine Solution 30-2214 L-glutamine is an essential amino acid required by virtually all 200mM(100 ml) mammalian and insect cells grown in culture.L-Alanyl-L-Glutamine Solution 30-2115 L-alanyl-L-glutamine can be used as a direct substitute for L-glu- 200mM(100 ml) tamine. Whereas L-glutamine is labile in cell culture mediumand its degradation results in deleterious build-up of ammonia,this dipeptide supplement is very stable with minimal ammoniaproduction.MEM Non-essential Amino 30-2116 This solution is added as a supplement to minimal basal media. Acid Solution (100 ml)The nonessential amino acids in this solution are 100 times the 100x concentration found in MEM-alpha Medium.Penicillin-Streptomycin-30-2220 This convenient solution allows researchers to add all these com- Glutamine Solution(100 ml) mon supplements in one step, saving time and reducing the riskof contamination.Penicillin-Streptomycin 30-2300 This solution reduces the chances of microbial contamination in Solution (100 ml) cell culture. Antibiotics may eliminate susceptible bacteria butcan also interfere with the metabolism of sensitive cells in culture. Water 30-2205 Meets WFI quality standards. For use as a diluent or forCell Culture Grade(500 ml) reconstituting powdered media and salt solutions.Hank’s Balanced Salt Solution30-2213 Contains no calcium, magnesium, or phenol red. For washing(500 ml) cells, f or use as a diluent, or for use as an inorganic base formedia preparation.I have been using ATCC media for over a year. I am verypleased with excellent cell growth, greater viability,and quick recover. ATCC high-performance media defi-nitely provide better results than other products I haveused in the past. I would highly recommend ATCC prod-ucts for any cell culture scientists in their daily works.—Shau-Ming Mong, Scientist, Ligand PharmaceuticalsATCC provides scientists worldwide with frozen vials of genuine, fully characterized and completely authenticated cell lines. Each bioproduction run of every cell line produced at ATCC is tested for growth and identity verification. The following tips are pro-vided by ATCC scientists to help ensure optimal results and less work overall.• Revive frozen cells and place them into cul-ture immediately upon receipt. If this is not possible, store cells in liquid nitrogen vapor (below –130º C).• Avoid “shocking” your cells. Most cell lines exhibit altered characteristics and need time to adapt when introduced to a new and different growth media. Using the same media in which ATCC cell lines were preserved will result in healthier, heartier cells and reduce cell loss due to adaptation.• Exercise patience. Cell lines react differently to cryopreservation and may require a few days in culture before they attach and grow.• Be cautious when purchasing media from alternative suppliers, as sodium bicarbonate concentrations may be higher than those em-ployed by ATCC. Slight changes in pH or reduced availability of dissolved CO2in the culture medium, may inhibit recovery of cells from cryopreservation or cause delayed attachment and slowed proliferation. All ATCC media, with the exception of Leibovitz’s L-15 (catalog no.30-2008), are formulated for use with 5% CO2.• Start with fresh cell lines. Cell lines that have been subcultured multiple times can experi-ence phenotypic and genotypic changes (ge-netic drift). If you start to experience sudden and inexplicable variations in your experimental results, it may be that the cell line has been sub-cultured too often and needs to be replaced.Over the course of more than 40 years, ATCC has generated a wealth of expertise in preserving and propagating cell lines. Contact ATCC Technical Ser-vice for further information.Phone 800-638-6597703-365-2700Fax 703-365-2750Email tech@After your expertise, the most important element of any experiment is the cell line Taking better care of your cellsP .O. Box 1549Manassas, VA 20108Phone: 800.638.6597 or 703.365.2700CB21-0906-02-02These products are for laboratory use only. Not for human or diagnostics use.© 2006 ATCC. All rights reserved. The ATCC trademark and trade name and any and all ATCC catalog numbers are trademarks ofFor more information or to order:Phone 800-638-6597 703-365-2700Fax 703-365-2750Emailsales@Visit us online at .。