【神经小胶质细胞】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。

【神经小胶质细胞】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%

【神经小胶质细胞】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

【神经小胶质细胞】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

4502103  artus VZV TM PCR Kit (24) RUO        CNY                                  

133271    NeXtalStock PIPES pH 7.0 (200)         CNY                                  

133270    NeXtalStock Phosphate-citr. pH 5.2 (200)           CNY                            【神经小胶质细胞】   

990512    Filter-Tips, 50 µl (960)         CNY                                  

337620    qBiomarker SYBR Green Fluor Mastermix(2)             CNY                                  

337011    qBiomarker Somatic Mutations PCR Assays        CNY                                  

4500263  artus HSV-1/2 RG PCR Kit (24) CE             CNY                                  

939016    Buffer ATL ( 4 x 50 ml),GPR        CNY                                  

4500163  artus HSV-1/2 TM PCR Kit (24) CE            CNY                                  

1073925  Ni-NTA Superflow (8 x 500 ml),cus,G        CNY                                  

9010931  BGR Greifer P7 (alt)             CNY                                  

9010858  BGR DT-Abstreifer 8-fach P7xxx D            CNY                                  

931036    QIAsymphony Virus/Bact Mini Kit (192)             CNY                                  

133248    NeXtalStock Sodium succinate pH6.5 (200)        CNY                                  

330603    RT² SYBR Green FAST Mastermix (24)              CNY                                  

929608    QX Pro Separation Buffer (40 ml)        CNY                                  

5000-1415      Knob RB671CSKWL Matte, VTX2              CNY                                  

5000-1408      Cable Shielded 1/17HP, VTX2              CNY                                  

L2170     hc2 CT/GC HC2 DNA Test PI (CD-ROM)          CNY                            【神经小胶质细胞】   

L1971     Rotary Shaker User＇s Manual        CNY                                  

9240705  O-ring Kit, Capper, Spindle, AP            CNY                                  

9240693  Actril Sterilant & Test Strips, AP          CNY                                  

9243653  Comprehen Valid., Other Extraction Sys       """The Comprehensive Validation Support plan specific to a laboratory＇s needs will incorporate all of the requirements for the internal validation guidelines from the FBI＇s Quality Assurance Standards (QAS).

The Comprehensive Validation Support service includes all travel ａnd labor charges for up to 20 days by the QIAGEN Validation Team. Typically, one week will be required to extract, quantify, ａnd amplify the validation study samples ａnd may require multiple individuals. Usually, two weeks is required to analyze the data ａnd one week to write the final report. The expected time period to complete the Comprehensive Validation is six weeks. The actual time may vary depending on resource availability, including laboratory time dedicated to validation activities.

Lab Work – The QIAGEN specialist(s) will travel to the laboratory to conduct the on-site validation lab work. The QIAGEN specialist team will perform ａnd complete all extractions, quantifications, amplfications, ａnd capillary electrophoresis. The laboratory is responsible for providing systems/protocols that may be needed (generally laboratory SOPs, ex. interpretation guidelines). Additionally, the laboratory staff may be required to provide additional data needed for interpretation (ex. quantification/amplification/CE results) during the data review phase of the validation.

Once the final scope of the validation is determined, QIAGEN will provide a quote for all consumables required for the validation.  Non-QIAGEN chemistry ａnd accessories are not included ａnd should  be purchased to cover the validation experimental design including qPCR, STR, NIST standards, required samples types (blood, semen, buccal samples) ａnd required substrates (swabs, materials, etc).   The Customer is expected to have all materials on-hand 1 week prior to the validation start date.

Data Analysis – The Comprehensive Validation Support plan encompasses data collection (quants, amps, CE) during the on-site visit. The laboratory may be responsible for sharing (typically electronically via e-mail) any validation data requested after the on-site visit. A QIAGEN specialist will complete all of the data analysis produced from the validation study design. This includes qPCR (quant) ａnd STR (amp) analysis ａnd linkage to starting samples.

Validation data CD/flashdrive – QIAGEN will review the validation data ａnd draft a validation summary document, ensuring compliance with the FBI＇s QAS requirements/standards. QIAGEN will send validation data in an electronic format (email/CD/flashdrive) to the laboratory. The laboratory is responsible for final compilation, any format change preferences, review ａnd signoff of the validation report. The documentation provided by QIAGEN will aide in demonstrating compliance with all accreditation guidelines during laboratory audits. Should a hard copy of the validation study be preferred, the laboratory is required to specify prior to the onset of the validation.

Post-validation QIAGEN-to-Laboratory Transfer – Following completion of the validation ａnd generation of the draft validation summary document, a designated member of the QIAGEN validation team will review the validation with the Lab Manager ａnd the Technical Leader. Following sign off on the validation report, training of the laboratory DNA staff will occur. Refer to the Post-validation Competency Training for additional information. """   CNY                                  

386516    Investigator STR GO! Lysis Buffer (200)            CNY                                  

208256    QuantiNova Probe PCR Kit (2500)              CNY                                  

208254    QuantiNova Probe PCR Kit (500)         CNY                                  

180984    GeneRead Adapter I Set 12plex (72)            CNY                                  

614035    careHPV Test, SFDA (96-test)             CNY                                  

282605    virotype Influenza A RT-PCR Kit (96)          CNY                                  

272703    pigtype PRRSV Ab (5)          CNY                                  

273003    pigtype Salmonella Ab (5)             CNY                                  

280377    virotype BVDV RT-PCR Kit (480)        CNY                                  

9018462  Marker Kit, QC tool, QX              CNY                                  

EK0398   mIL-2-ELISA 小鼠IL-2ELISA试剂盒       Mouse IL-2 ELISA Kit   96 tests                

EK0403   m IL-3 -ELISA      小鼠IL-3ELISA试剂盒       Mouse IL-3 ELISA Kit   96 tests                 【神经小胶质细胞】

EK0405   m IL-4-ELISA       小鼠IL-4 ELISA试剂盒      Mouse IL-4 ELISA Kit   96 tests                

EK0408   m IL-5-ELISA       小鼠IL-5ELISA试剂盒       Mouse IL-5 ELISA Kit   96 tests                

EK0411   m IL-6-ELISA       小鼠IL-6ELISA试剂盒       Mouse IL-6 ELISA Kit   96 tests                

EK0417   mIL-10-ELISA       小鼠IL-10ELISA试剂盒      Mouse IL-10 ELISA Kit 96 tests                

EK0422   mIL-12(p70)-ELISA      小鼠IL-12(p70)ELISA试剂盒     Mouse IL-12(p70) ELISA Kit 96 tests                

EK0431   mIL-17-ELISA       小鼠IL-17ELISA试剂盒      Mouse IL-17 ELISA Kit 96 tests                

EK0436   mLaminin-ELISA   小鼠层粘连蛋白ELISA试剂盒   Mouse Laminin ELISA Kit     96 tests                

EK0438   mLeptin-ELISA      小鼠瘦蛋白ELISA试剂盒   Mouse Leptin ELISA Kit 96 tests                

EK0440   mLeptinR-ELISA    小鼠瘦蛋白受体ELISA试剂盒   Mouse Leptin receptor ELISA Kit  96 tests                

EK0445   mMCSF-ELISA     小鼠巨噬细胞集落刺激因子ELISA试剂盒       Mouse M-CSF ELISA Kit      96 tests                

EK0460   mMMP-2-ELISA    小鼠MMP-2ELISA试剂盒   Mouse MMP-2 ELISA Kit     96 tests                

EK0462   mMMP3-ELISA     小鼠MMP-3ELISA试剂盒   Mouse MMP-3 ELISA Kit     96 tests                

EK0466   mMMP-9-ELISA    小鼠MMP-9ELISA试剂盒   Mouse MMP-9 ELISA Kit     96 tests                

EK0470   mNGF-ELISA 小鼠NGF/NGFβELISA试剂盒  Mouse NGF/NGFβ ELISA Kit    96 tests                

EK0473   mNT3-ELISA 小鼠Neurotrophin-3 ELISA试剂盒     Mouse Neurotrophin-3 ELISA Kit  96 tests                

EK0483   mOPN-ELISA 小鼠骨桥素Elisa试剂盒ELISA试剂盒      Mouse OPN ELISA Kit  96 tests                 【神经小胶质细胞】

EK0486   mPDGF-AB-ELISA       小鼠血小板衍生生长因子ABELISA试剂盒      Mouse PDGF-AB ELISA Kit  96 tests                

EK0495   mRantes-ELISA     小鼠趋化因子ELISA试剂盒       Mouse Rantes ELISA Kit       96 tests                

EK0504   mSELL-ELISA       小鼠L选择素Elisa试剂盒   Mouse L-Selectin ELISA Kit  96 tests                

EK0506   mSELP-ELISA       小鼠P选择素Elisa试剂盒   Mouse P-Selectin ELISA Kit  96 tests                

EK0515   mTGF     小鼠转化生长因子β1ELISA试剂盒  Mouse TGFβ1 ELISA Kit    96 tests                

EK0527   mTNF-ELISA 小鼠TNFαELISA试剂盒   Mouse TNFα ELISA Kit     96 tests                

EK0538   mVCAM-ELISA     小鼠血管细胞粘附分子-1ELISA试剂盒    Mouse VCAM-1 ELISA Kit    96 tests                

EK0541   mVEGF-ELISA      小鼠血管内皮生长因子ELISA试剂盒       Mouse VEGF ELISA Kit 96 tests                

EK0562   mECad-ELISA       小鼠钙粘着蛋白ELISA试剂盒   Mouse E-Cadherin ELISA Kit 96 tests                

EK0577   mGp130-ELISA     小鼠Gp130/IL6STELISA试剂盒 Mouse Gp130/IL6ST ELISA Kit    96 tests                

EK0582   mRes-ELISA  小鼠抵抗素ELISA试剂盒   Mouse Resistin ELISA Kit     96 tests                

EK0664   mBAFF-ELISA      小鼠BAFFELISA试剂盒     Mouse BAFF ELISA Kit 96 tests                

EK0668   mPCad-ELISA       小鼠P-CadherinELISA试剂盒     Mouse P-Cadherin ELISA Kit 96 tests