【肾皮层上皮细胞】下面是细胞株培养的具体无菌技术要点：

1. 实验开始前，无菌室和无菌操作台需要紫外光照射30到60分钟完全灭菌，用70%ethanol擦拭无菌操作台面，并且打开无菌操作台风扇运行10分钟之后，才可以进行实验操作。每次操作只能处理一株细胞株，即使培养基完全相同也不可共用培养基，以避免细胞间污染或失误混淆。实验完成后，请将实验物品拿出工作台，用70%ethanol再次擦拭无菌操作台面。操作间隔应该让无菌操作台运转10分钟到15分钟后，才能进行下一个细胞株的操作。

2. 无菌操作工作的区域应保持清洁和宽敞，必要物品（试管架、吸管、吸取器或吸管盒等）可以暂时放置，其它实验用品使用完毕后应移出，有利于空气流通。实验用品需70%ethanol擦拭后方可带入无菌操作台内。实验操作过程应在台面的中央无菌区域内完成，请勿在边缘非无菌区域进行操作。

3. 小心取用无菌实验物品，避免细菌污染。请勿触碰吸管尖头部和容器瓶口处，也不要在打开的容器正上方进行操作。容器打开后，请用手夹住瓶盖并轻握瓶身，倾斜大约45°角取用，尽量不要将瓶盖口朝上放置于桌面。

4. 工作人员应当首先注意自身安全，必须穿戴齐实验衣和实验手套后才能进行实验。对于病毒感染或是来自人类的细胞株应当特别小心操作，并选择适当等级的无菌操作台进行（至少是Class II）。操作过程中，应避免引起aerosol的产生，小心有毒性药品，例如DMSO和TPA等，并避免针头的伤害等等。

5. 定期检测下列项目：

5.1. CO2钢瓶的CO2压力

5.2. CO2培养箱的CO2浓度、温度、和水盘是否有污染（水盘的水需用无菌水，每周定时更换）。

5.3. 无菌操作台内的airflow压力，定期更换紫外线灯管和HEPA过滤膜，预滤网（300小时/预滤网，3000小时/HEPA）。

6. 水槽内可添加消毒剂（Zephrin 1:750），需定期更换水槽中的水。

【肾皮层上皮细胞】齐一生物友情提醒，本产品仅供科研使用，不得用于临床及诊断使用！

一细胞说明，具体详细请见细胞附带说明书

 二、客户自备试剂

1、PBS  

2、 2、RPMI-1640 培养基（不含 Hepes）+ 10%胎牛血清+ 1%双抗

3、0.25%  （W/V）胰蛋白酶（含 0.02% EDTA）

三、细胞背景

1、生长方式：贴壁

2、种属：Homo sapiens  人

四、培养条件

1、培养基：RPMI-1640 培养基（不含 Hepes）+ 10%胎牛血清+ 1%双抗

2、温度：37.0°C

3、气体:空气  95%，CO2 5%

【肾皮层上皮细胞】五、培养方法

 收到细胞后，在倒置显微镜下观察整个细胞生长情况：

 如果细胞未长满，用 75%酒精喷洒整个瓶消毒后放到超菌台内，严格无菌操作，打开细胞培养瓶，留10ml 培养液继续培养。如果细胞已长满（达 80-90%）。即可进行传代，具六、体步骤如下：

1，弃去培养液，用PBS洗 1-2次。

2，向瓶内加入 1.0-2.0ml 胰蛋白酶液，在倒置显微镜下观察细胞消化情况，若细胞大部分变圆，迅速拿回操作台，吸取胰蛋白酶，加含有 6ml  含 10%血清的培养液，轻轻吹打细胞。

3，加入等量的的培养液，轻轻吹打混匀后吸出一半，分到新的培养

4，传代比例：1:2-1:3  

七、常见问题及解决方案

1、培养瓶有破裂，培养液有漏液：细胞极大可能会污染，所以我们会及时安排帮老师解决。

2、细胞漂浮：培养瓶不开封，瓶口酒精擦拭后平躺放置在培养箱。次日观察，如细胞大部分又贴回瓶底，表明细胞活力正常，剩余漂浮的细胞可以去掉，留 10ml 培养液培养观察，细胞生长至汇合度 80%，进行消化传代；如细胞还是不贴壁，将细胞离心收集转到新培养瓶，原培养瓶加部分培养液继续培养，中间注意观察，我们的技术人员会一直跟踪指导，直到问题解决。

细胞培养代理品牌

细胞培养代理品牌

【肾皮层上皮细胞】齐一生物科技（上海）有限公司是一家专业生产和销售各种生化试剂,细胞株、原代细胞、菌种、医药中间体,标准品和对照品的大型化学科技公司. 齐一生物销售：021－6034 8496；181214 53965；173021 04490；Web:www.qiyibio.com

9235161  Zylinderstift rostfrei 2 h8 x25              CNY                                  

9230253  Rack for reagents, Universal (zig-zag)         CNY                                  

9022779  PyroMark Q24 Advanced Software             CNY                                  

9235105  Manual, user, hardware BR8000 Int             CNY                                  

9234863  Washer liquid container (9l Nalgene)            CNY                                  

9238670  Wedge belt 5 x 513        CNY                                  

9238648  Motorized lock with optic, right    "For Sigma centrifuge

Motor that controls the locks

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9238640  Tachogenerator             CNY                                  

9242058  Tube insert, 13 mm, R, sample carr., Qsy           CNY                                  

9241809  Pyro Q96 MDa, Installation   Installation ａnd Introductory Training PyroMark is an on-site service, conducted by a QIAGEN Service Specialist. It covers the installation ａnd commissioning of the instrument ａnd the handling of the PyroMark systems hardware. It trains the user on the operation of the PyroMark software ａnd the setup ａnd execution of a pyrosequencing run. After the training the user will be able to prepare samples, set-up ａnd run the instrument, analyse the results ａnd design new assays as well as troubleshoot new assays ａnd perform basic maintenance on the sample preparation tool ａnd instrument. All travel ａnd labor costs ａnd well as material for the demonstration run are included. The service is documented in accordance with GMP/GLP requirements       CNY                                  

9241162  Board, transport conversion, QX          CNY                                  

9241146  RS232 Board, Eppendorf, AP196/98            CNY                                  

9240993  Calibration Plate, MDD         CNY                                  

9240962  PCB, connector, pipetting system, Qsym            CNY                                  

9240959  Activation bar tip, handler, Qsym         CNY                            【肾皮层上皮细胞】   

9240948  Cable, x, portal, Qsym          CNY                                  

9240825  Field Upgrade, BR EZ1 DSP         CNY                                  

9240817  #Premium Cover Agreement EZ1 DSP, 1YR "Repair service delivery: onsite

Service response time (repair): 24 working hours/1 business day

Periodic inspection/maintenance: yes, 1/agreement year

Inspection/maintenance delivery: onsite

Cost coverage for repair parts: yes, included

Cost coverage for labor: yes, included

Cost coverage for travel: yes, included

Replacement system (Loaner) provision: not included

Transportation cost coverage: not applicable"      CNY                                  

9240776  Dispense Head Update, AP96        CNY                                  

9022763  EZ1 Adv. XL DNA Invest. Flip-Cap Card            CNY                                  

6000-5012      Lumicheck Battery Replacement Package            CNY                                  

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9241774  QIAgility, Installation     The Installation ａnd Introductory Training is an on-site service, conducted by a QIAGEN Service Specialist. It covers the handling of the QIAgility systems hardware ａnd trains the user on the operation of the software. After the training the user will be able to run ａnd install application programs ａnd conduct a basic troubleshooting. All travel ａnd labor costs are included. The service is documented in accordance with GMP/GLP requirements.   CNY                                  

9241655  Input Rack Pins Kit, AP        CNY                                  

9241360  Plastic supporters, door hinge, EZ1a            CNY                                  

9243547  Pyro Q24, PM Service   "

Service response time (repair): not applicable

Periodic inspection/maintenance: yes

Inspection/maintenance delivery: onsite

Cost coverage for Repair Parts : not included

Cost coverage for labor: yes, full coverage

Cost coverage for travel: yes, full coverage

Replacement system (Loaner) provision: not applicable

Transportation cost coverage: yes, full coverage" CNY                                  

9243112  Hard Drive, Programmed, Cheetah, AP98            CNY                                  

9242770  Belt, x-drive, portal, AS, Qsym            CNY                                  

9242698  RGQ, Full Agreement     "Repair service delivery: send to Regional Repair Center

Service response time (repair): 7-10 days turnaround time

Periodic inspection/maintenance: yes, 1/agreement year

Inspection/maintenance delivery: onsite

Cost coverage for repair parts: yes, included

Cost coverage for labor: yes, included

Cost coverage for travel: yes, included

Replacement system (Loaner) provision: yes, included (provision time 2 days)

Transportation cost coverage: yes, included"       CNY                                  

5000-1406      Control PCB 120V, VTX2             CNY                                  

5000-1320      Label Front Overlay, SKR1           CNY                            【肾皮层上皮细胞】   

5000-1312      Platform, SKR1             CNY                                  

5000-1274      Plate Mask, DML2        CNY                                  

QTR-981 马传染性动脉炎iELISA抗体检测试剂盒                                    

QTR-982 马疱疹病毒抗体检测试剂盒                                         

QTR-983 西尼罗河热IgM抗体检测试剂盒                                         

QTR-984 兔粘液瘤病毒抗体检测试剂盒                                      

QTR-985 兔出血病病毒抗体检测试剂盒                                      

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QTR-987 伯氏疏螺旋体抗体检测试纸条                                      

QTR-988 犬冠状病毒抗体检测试剂盒                                         

QTR-989 犬瘟热IgG检测试剂盒                                         

QTR-990 犬瘟热IgM检测试剂盒                                         

QTR-991 犬细小IgM检测试剂盒                                         

QTR-992 犬细小抗体检测试剂盒                                         

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QTR-997 犬心丝虫抗原检测试纸条                                      

QTR-998 犬利什曼抗体检测试剂盒                                      

QTR-999 犬利什曼抗体检测试纸条                                      

QTR-1000      狂犬病抗原检测试剂盒                                         

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