【虹膜色素上皮细胞总DNA】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。

【虹膜色素上皮细胞总DNA】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%

【虹膜色素上皮细胞总DNA】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

【虹膜色素上皮细胞总DNA】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

9236417  Software, Twister I, TWI            CNY                                  

9236264  Elution Microtubes Adaptor, inject mold              CNY                                  

9236165  O-ring, lubrication kit, RapidPlate         CNY                                  

9235931  Splashing guard, BR8 RD             CNY                                  

9230822  Rack for reagents,Trough (for 6 troughs)           CNY                                  

9235707  Washer Cable, RCS1             CNY                                  

9235695  Ground Wire, RCS1              CNY                                  

9235332  Z-Rack Packing Cover          CNY                                  

9235118  Fuse F1A, v1.00           CNY                                  

9235114  Fuse FST4A, v1.00              CNY                                  

9230054  Clamp, Tubing, Z-bar           CNY                                  

9234861  Lid coupling piece for system liquid             CNY                                  

9239551  Cbl Assy, XY, LED Driver            CNY                                  

9238663  Hook, motorized lid lock              CNY                                  

9238642  Capacitor board             CNY                                  

9238626  Opto-Encoder        CNY                            【虹膜色素上皮细胞总DNA】  

9238625  Eprom ａnd controller 4K15, Zent3C            CNY                                  

9241828  Pyro Q96 MDa, Premium Agreement    "Repair service delivery: onsite

Service response time (repair): 24 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                                  

9241122  Superlube              CNY                                  

9241007  QIArent&go - Rent out fee QIAcube           CNY                                  

9240985  PCB, drive, cooling device, Qsym        CNY                                  

9240982  Sensor, x, sample tracking, Qsym        CNY                                  

9240951  Support, cable, portal, Qsym        CNY                                  

9234820  Manual, user, QS 4.0, Int             CNY                                  

9240768  Signal Conditioner, Centrifuge, AP96           CNY                                  

9240764  DC Output Module 8NPT, 3AMP, AP96              CNY                                  

6000-5003      Duraseal Sealing Film            CNY                                  

9241781  RGQ, Full Agreement, no PM       "Repair service delivery: send to Regional Repair Center

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

Periodic inspection/maintenance: not included

Inspection/maintenance delivery: not applicable

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                                  

9241776  QIAgility, Inspection Service "The Annual Inspection Service, QIAgility is an on-site equipment service, performed by a QIAGEN service specialist.

All required testing material ａnd certified equipment is provided by the service specialist.

During the half day-long procedure, all application-critical modules of the QIAgility are inspected ａnd tested ａnd a pipetting head calibration is conducted. Test results are documented in a GMP/GLP-compliant Visit Report." CNY                                  

9241756  IQ/OQ Services, QIAsymphony    Instrument qualification support service for the QIAsymphony. The product contains the Installation Qualification (IQ)/Operation Qualification (OQ) documentation ａnd the performance of the qualification protocols (travel ａnd labor costs included).      CNY                                  

9237126  Upgrade App., PCR Setup            CNY                                  

9237093  Training, Extension day  Additional days of training may be purchased during the week of the site visit upon request.       CNY                            【虹膜色素上皮细胞总DNA】  

9236426  Board, arm interconnect, TWI             CNY                                  

QY-x1172       人乳腺导管癌细胞MDA-MB-453                                         

QY-x1173       人乳腺细胞HBL-100                                      

QY-x1174       人乳腺癌细胞BCaP-37                                   

QY-x1175       人乳腺癌细胞LCC1                                       

QY-x1176       人乳腺癌细胞MX-1                                       

QY-x1177       人乳腺管癌细胞T47D                                    

QY-x1178       人乳腺癌细胞SK-BR-3                                    【虹膜色素上皮细胞总DNA】

QY-x1179       人乳腺癌细胞4T1                                   

QY-x1180       人乳腺癌细胞HCC1937                                         

QY-x1181       人乳腺导管癌细胞HCC38                                     

QY-x1182       人乳腺癌细胞Hs-578T                                   

QY-x1183       人乳腺癌Tamoxifen耐药株LCC2                                        

QY-x1184       人乳腺癌抗雌激素耐药株LCC9                                    

QY-x1185       人乳腺癌阿霉素耐药细胞株MCF/Adr                                         

QY-x1186       人乳腺癌细胞BT-549                                     

QY-x1187       人乳腺导管瘤细胞BT-474                                     

QY-x1188       人乳腺癌细胞MB-271                                    

QY-x1189       人乳腺癌细胞ER-30                                      

QY-x1190       人乳腺癌细胞M453                                       

QY-x1191       人乳腺浸润性导管癌旁皮肤细胞CCD-1095Sk                                   

QY-x1192       人乳腺细胞HBL-100                                      

QY-x1193       人乳腺导管瘤细胞UACC812                                        

QY-x1194       人乳腺癌细胞系1590                                      【虹膜色素上皮细胞总DNA】

QY-x1195       人卵巢癌细胞A2780                                      

QY-x1196       人卵巢癌细胞Sw626                                     

QY-x1197       人卵巢癌顺铂耐药株COC1/CDDP                                       

QY-x1198       人卵巢癌紫杉醇耐药株A2780/Taxol                                    

QY-x1199       人卵巢癌细胞系Anglne                                         

QY-x1200       人卵巢癌细胞OVCaR-3                                         

QY-x1201       人卵巢癌细胞Caov-3                                     

QY-x1202       人卵巢癌细胞HO-8910