【小腸粘膜上皮細胞】下面是細胞株培養的具體無菌技術要點：

1. 實驗開始前，無菌室和無菌操作臺需要紫外光照射30到60分鐘完全滅菌，用70%ethanol擦拭無菌操作臺面，并且打開無菌操作臺風扇運行10分鐘之后，才可以進行實驗操作。每次操作只能處理一株細胞株，即使培養基完全相同也不可共用培養基，以避免細胞間污染或失誤混淆。實驗完成后，請將實驗物品拿出工作臺，用70%ethanol再次擦拭無菌操作臺面。操作間隔應該讓無菌操作臺運轉10分鐘到15分鐘后，才能進行下一個細胞株的操作。

2. 無菌操作工作的區域應保持清潔和寬敞，必要物品（試管架、吸管、吸取器或吸管盒等）可以暫時放置，其它實驗用品使用完畢后應移出，有利于空氣流通。實驗用品需70%ethanol擦拭后方可帶入無菌操作臺內。實驗操作過程應在臺面的中央無菌區域內完成，請勿在邊緣非無菌區域進行操作。

3. 小心取用無菌實驗物品，避免細菌污染。請勿觸碰吸管尖頭部和容器瓶口處，也不要在打開的容器正上方進行操作。容器打開后，請用手夾住瓶蓋并輕握瓶身，傾斜大約45°角取用，盡量不要將瓶蓋口朝上放置于桌面。

4. 工作人員應當首先注意自身安全，必須穿戴齊實驗衣和實驗手套后才能進行實驗。對于病毒感染或是來自人類的細胞株應當特別小心操作，并選擇適當等級的無菌操作臺進行（至少是Class II）。操作過程中，應避免引起aerosol的產生，小心有毒性藥品，例如DMSO和TPA等，并避免針頭的傷害等等。

5. 定期檢測下列項目：

5.1. CO2鋼瓶的CO2壓力

5.2. CO2培養箱的CO2濃度、溫度、和水盤是否有污染（水盤的水需用無菌水，每周定時更換）。

5.3. 無菌操作臺內的airflow壓力，定期更換紫外線燈管和HEPA過濾膜，預濾網（300小時/預濾網，3000小時/HEPA）。

6. 水槽內可添加消毒劑（Zephrin 1:750），需定期更換水槽中的水。

【小腸粘膜上皮細胞】齊一生物友情提醒，本產品僅供科研使用，不得用于臨床及診斷使用！

一細胞說明，具體詳細請見細胞附帶說明書

 二、客戶自備試劑

1、PBS  

2、 2、RPMI-1640 培養基（不含 Hepes）+ 10%胎牛血清+ 1%雙抗

3、0.25%  （W/V）胰蛋白酶（含 0.02% EDTA）

三、細胞背景

1、生長方式：貼壁

2、種屬：Homo sapiens  人

四、培養條件

1、培養基：RPMI-1640 培養基（不含 Hepes）+ 10%胎牛血清+ 1%雙抗

2、溫度：37.0°C

3、氣體:空氣  95%，CO2 5%

【小腸粘膜上皮細胞】五、培養方法

 收到細胞后，在倒置顯微鏡下觀察整個細胞生長情況：

 如果細胞未長滿，用 75%酒精噴灑整個瓶消毒后放到超菌臺內，嚴格無菌操作，打開細胞培養瓶，留10ml 培養液繼續培養。如果細胞已長滿（達 80-90%）。即可進行傳代，具六、體步驟如下：

1，棄去培養液，用PBS洗 1-2次。

2，向瓶內加入 1.0-2.0ml 胰蛋白酶液，在倒置顯微鏡下觀察細胞消化情況，若細胞大部分變圓，迅速拿回操作臺，吸取胰蛋白酶，加含有 6ml  含 10%血清的培養液，輕輕吹打細胞。

3，加入等量的的培養液，輕輕吹打混勻后吸出一半，分到新的培養

4，傳代比例：1:2-1:3  

七、常見問題及解決方案

1、培養瓶有破裂，培養液有漏液：細胞極大可能會污染，所以我們會及時安排幫老師解決。

2、細胞漂�。号囵B瓶不開封，瓶口酒精擦拭后平躺放置在培養箱。次日觀察，如細胞大部分又貼回瓶底，表明細胞活力正常，剩余漂浮的細胞可以去掉，留 10ml 培養液培養觀察，細胞生長至匯合度 80%，進行消化傳代；如細胞還是不貼壁，將細胞離心收集轉到新培養瓶，原培養瓶加部分培養液繼續培養，中間注意觀察，我們的技術人員會一直跟蹤指導，直到問題解決。

細胞培養代理品牌

細胞培養代理品牌

【小腸粘膜上皮細胞】齊一生物科技（上海）有限公司是一家專業生產和銷售各種生化試劑,細胞株、原代細胞、菌種、醫藥中間體,標準品和對照品的大型化學科技公司. 齊一生物銷售：021－6034 8496；181214 53965；173021 04490；Web:www.qiyibio.com

9230281  Cable, Interface, Plato-Syringe Pump          CNY                                  

9235253  Trough-Rack spezial             CNY                                  

9230197  Plug f. dilutor valve inlet        CNY                            【小腸粘膜上皮細胞】      

9230059  Rack for reagents, Trough(for 3 troughs)           CNY                                  

9238674  Bracket for grey wedge         CNY                                  

9238652  Interference suppression capacitor              CNY                                  

9238636  Gear motor, lid opening         CNY                                  

9242027  Repair fee, Depot Repair, Palm Cycler         CNY                                  

9241822  Pyro Q24, 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

 or Repair Center Cost coverage for Repair Parts : yes, full coverage

Cost coverage for labor: yes, full coverage

Cost coverage for travel: yes, full coverage

Replacement system (Loaner) provision: yes, included (provision time 2-3 days)

Transportation cost coverage: yes, full coverage"""     CNY                                  

9241819  Pyro Q96 ID, Basic Agreement     "Repair service delivery: onsite

Service response time (repair): 5 business days

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                                  

9240975  PCB, control, sample tracking, Qsym          CNY                                  

9240969  PCB, carriersens, sample tracking, Qsym            CNY                                  

9240682  Dump Tray, Overflow Control, AP98          CNY                                  

9240789  Update,Unload Valve Assy, AP196/98           CNY                                  

9240815  #Basic Cover Agreement EZ1 DSP, 1YR             CNY                                  

9240775  Sample Rack, AP           CNY                                  

6000-5017      MST Conv.Rack&Lid (Silver - 2 pos. cal.)          CNY                                  

9241762  QIAgility, Basic Agreement    "Repair service delivery: onsite

Service response time (repair): 5 business days

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) or PC repair provision: not included

Transportation cost coverage: not applicable"      CNY                                  

9241380  24VDC Adaptor Cable, Microlynx, AP         CNY                            【小腸粘膜上皮細胞】      

9240758  Update, Bracket, Shaker Bimba, AP98          CNY                                  

9243552  Target Prep, 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                                  

9243384  Cooling adapter, SBS Universal, Qsym         CNY                                  

9243116  Lee Sample Valve Kit, LC             CNY                                  

9243234  HC2, PM Agreement      The Preventive Maintenance (PM) service ensures the correct operation of the equipment. During the PM, all the instrument modules are inspected by a qualified QIAGEN service technician. The Preventive Maintenance is performed under the manufacturer guidelines ａnd documented according to GLP / GMP   CNY                                  

9243044  Set, brackets, transparent panel, Qsym        CNY                                  

9242752  Kit, upg-x/y-drive, portal, Qsym          CNY                                  

9242711  Validation Support, 2d, QIAsymphony  The QIAGEN QIAsymphony Guided Validation Support service includes all travel ａnd labor charges for a site visit up to 2 days by a QIAGEN Applications Specialist.  A QIAGEN specialist will provide an introduction to the QIAsymphony system ａnd the DNA Investigator silica chemistry utilized in the process, review the Handbook ａnd User Manual, as well as hands-on QIAsymphony instrument operation.  The Guided Validation service includes experimental design consultation, internal validation documentation support, ａnd QIAGEN sponsored data templates. The QIAGEN Specialist will also aide the customer in running some samples to initiate the validation process.  Enough consumable chemistry ａnd accessories (QIAGEN or other vendors) should be purchased by the customer to cover the validation experimental design ａnd are not included in the cost of validation support. Please indicate which protocols to include in the validation (Casework200, Casework500, Reference200, etc.).      CNY                                  

9242203  Cooling adapter, SBS Universal, Qsym         CNY                                  

9242198  Lid Sealing Ring, Eppendorf, AP196/98        CNY                                  

5000-1402      Latch Front Assembly, VTX2              CNY                                  

5000-1315      Rotary Shaker - Spring Single              CNY                                  

5000-1308      Sensor Speed, SKR1             CNY                                  

5000-1280      Bottom Feet W Screws, DML2            CNY                                  

QTR-207 痢疾2型診斷血清       1ml/瓶                             

QTR-208 宋內氏1相診斷血清    1ml/瓶                              【小腸粘膜上皮細胞】

QTR-209 宋內氏2相診斷血清    1ml/瓶                             

3、大腸艾希氏菌診斷血清                                           

QTR-301 致病性大腸艾希氏菌診斷血清15種  1ml×18瓶                              

QTR-302 侵襲性大腸艾希氏菌診斷血清11種  1ml×11瓶        供研究用                      

QTR-303 產毒性大腸艾希氏菌診斷血清10種  1ml×10瓶        供研究用                      

QTR-304 出血性大腸艾希氏菌O157診斷血清 1ml/瓶       供研究用                      

QTR-305 大腸艾希氏菌H7診斷血清  1ml/瓶       供研究用                      

4、霍亂弧菌診斷血清                                                      

QTR-401 O1群霍亂弧菌診斷血清3種      1ml×11瓶                                

QTR-402 O1群、O139霍亂弧菌多價診斷血清 1ml/瓶    供研究用                           

QTR-403 O139霍亂弧菌多價診斷血清      1ml/瓶    供研究用                           

5、診斷菌液系列                                           

QTR-501 傷寒、副傷寒及變形桿菌診斷菌液         10ml×5瓶      肥達氏                         

QTR-502 傷寒、副傷寒及變形桿菌診斷菌液         10ml×3瓶   外斐氏                        

QTR-503 傷寒、副傷寒及變形桿菌診斷菌液         10ml×8瓶     肥達、外斐                           

QTR-504 傷寒、副傷寒及變形桿菌診斷菌液(質控)      10ml×5瓶     配質控血清                            【小腸粘膜上皮細胞】

QTR-505 傷寒、副傷寒及變形桿菌診斷菌液(質控)      10ml×3瓶     配質控血清                           

6、特免血漿篩選試劑系列（ELISA）                                               

QTR-601 人狂犬病病毒IgG抗體測定試劑盒    96T                           

QTR-602 破傷風IgG抗體測定試劑盒       96T   孔內稀釋                        

QTR-603 乙型肝炎病毒表面IgG抗體測定試劑盒    96T   孔內稀釋                        

7、疫苗免疫效果監測試劑系列（ELISA）                                               

QTR-701 人狂犬病病毒IgG抗體測定試劑盒    48T                                 

QTR-702 人狂犬病病毒IgG抗體測定試劑盒    96T                           

QTR-703 動物狂犬病病毒IgG抗體測定試劑盒       48T   供研究用                        

QTR-704 破傷風IgG抗體測定試劑盒       48T   供研究用                        

QTR-705 出血熱IgG抗體測定試劑盒       48T   供研究用                        

QTR-706 風疹病毒IgG抗體測定試劑盒    48T    供研究用                      

QTR-707 麻疹病毒IgG抗體測定試劑盒    48T    供研究用