【心肌細(xì)胞總DNA】下面是細(xì)胞株培養(yǎng)的具體無菌技術(shù)要點(diǎn)：

1. 實(shí)驗(yàn)開始前，無菌室和無菌操作臺(tái)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(tái)面，并且打開無菌操作臺(tái)風(fēng)扇運(yùn)行10分鐘之后，才可以進(jìn)行實(shí)驗(yàn)操作。每次操作只能處理一株細(xì)胞株，即使培養(yǎng)基完全相同也不可共用培養(yǎng)基，以避免細(xì)胞間污染或失誤混淆。實(shí)驗(yàn)完成后，請(qǐng)將實(shí)驗(yàn)物品拿出工作臺(tái)，用70%ethanol再次擦拭無菌操作臺(tái)面。操作間隔應(yīng)該讓無菌操作臺(tái)運(yùn)轉(zhuǎn)10分鐘到15分鐘后，才能進(jìn)行下一個(gè)細(xì)胞株的操作。

2. 無菌操作工作的區(qū)域應(yīng)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(shí)放置，其它實(shí)驗(yàn)用品使用完畢后應(yīng)移出，有利于空氣流通。實(shí)驗(yàn)用品需70%ethanol擦拭后方可帶入無菌操作臺(tái)內(nèi)。實(shí)驗(yàn)操作過程應(yīng)在臺(tái)面的中央無菌區(qū)域內(nèi)完成，請(qǐng)勿在邊緣非無菌區(qū)域進(jìn)行操作。

3. 小心取用無菌實(shí)驗(yàn)物品，避免細(xì)菌污染。請(qǐng)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(jìn)行操作。容器打開后，請(qǐng)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(yīng)當(dāng)首先注意自身安全，必須穿戴齊實(shí)驗(yàn)衣和實(shí)驗(yàn)手套后才能進(jìn)行實(shí)驗(yàn)。對(duì)于病毒感染或是來自人類的細(xì)胞株應(yīng)當(dāng)特別小心操作，并選擇適當(dāng)?shù)燃?jí)的無菌操作臺(tái)進(jìn)行（至少是Class II）。操作過程中，應(yīng)避免引起aerosol的產(chǎn)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(cè)下列項(xiàng)目：

5.1. CO2鋼瓶的CO2壓力

5.2. CO2培養(yǎng)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(shí)更換）。

5.3. 無菌操作臺(tái)內(nèi)的airflow壓力，定期更換紫外線燈管和HEPA過濾膜，預(yù)濾網(wǎng)（300小時(shí)/預(yù)濾網(wǎng)，3000小時(shí)/HEPA）。

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

【心肌細(xì)胞總DNA】齊一生物友情提醒，本產(chǎn)品僅供科研使用，不得用于臨床及診斷使用！

一細(xì)胞說明，具體詳細(xì)請(qǐng)見細(xì)胞附帶說明書

 二、客戶自備試劑

1、PBS  

2、 2、RPMI-1640 培養(yǎng)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(xì)胞背景

1、生長(zhǎng)方式：貼壁

2、種屬：Homo sapiens  人

四、培養(yǎng)條件

1、培養(yǎng)基：RPMI-1640 培養(yǎng)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%

【心肌細(xì)胞總DNA】五、培養(yǎng)方法

 收到細(xì)胞后，在倒置顯微鏡下觀察整個(gè)細(xì)胞生長(zhǎng)情況：

 如果細(xì)胞未長(zhǎng)滿，用 75%酒精噴灑整個(gè)瓶消毒后放到超菌臺(tái)內(nèi)，嚴(yán)格無菌操作，打開細(xì)胞培養(yǎng)瓶，留10ml 培養(yǎng)液繼續(xù)培養(yǎng)。如果細(xì)胞已長(zhǎng)滿（達(dá) 80-90%）。即可進(jìn)行傳代，具六、體步驟如下：

1，棄去培養(yǎng)液，用PBS洗 1-2次。

2，向瓶?jī)?nèi)加入 1.0-2.0ml 胰蛋白酶液，在倒置顯微鏡下觀察細(xì)胞消化情況，若細(xì)胞大部分變圓，迅速拿回操作臺(tái)，吸取胰蛋白酶，加含有 6ml  含 10%血清的培養(yǎng)液，輕輕吹打細(xì)胞。

3，加入等量的的培養(yǎng)液，輕輕吹打混勻后吸出一半，分到新的培養(yǎng)

4，傳代比例：1:2-1:3  

七、常見問題及解決方案

1、培養(yǎng)瓶有破裂，培養(yǎng)液有漏液：細(xì)胞極大可能會(huì)污染，所以我們會(huì)及時(shí)安排幫老師解決。

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

細(xì)胞培養(yǎng)代理品牌

細(xì)胞培養(yǎng)代理品牌

【心肌細(xì)胞總DNA】齊一生物科技（上海）有限公司是一家專業(yè)生產(chǎn)和銷售各種生化試劑,細(xì)胞株、原代細(xì)胞、菌種、醫(yī)藥中間體,標(biāo)準(zhǔn)品和對(duì)照品的大型化學(xué)科技公司. 齊一生物銷售：021－6034 8496；181214 53965；173021 04490；Web:www.qiyibio.com

9236476  #Agreement, 9604, 2PM, 1 yr       An agreement for preventive maintenance provides coverage for travel, labor, ａnd parts, plus the cost of consumables required for two maintenance visits during 1 year. Work carried out during the preventive maintenance visit is fully documented in accordance with GMP/GLP directives.    CNY                                  

9232726  Block, 2-way, v1.02             CNY                                  

9239540  Cable Assy Extender, 2 wire, 22AWG, 12in          CNY                                  

9239396  Upgr. LiquiChip Analyzer (RUO)          CNY                                  

9239496  EX XY Motor Driver w/Encoder          CNY                            【心肌細(xì)胞總DNA】  

9239494  Sheath Fluid Tubing w/DeBubbler        CNY                                  

9239487  EX Analog PCBA           CNY                                  

9236515  Centrifuge, 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                                  

9232888  Camera, bar-code, BL180, P3xxx/BR3000          CNY                                  

9236636  Cover, Wash Station, BR8 RD              CNY                                  

9236451  Travel Charge, additional, 1 day           CNY                                  

9232725  Fitting, pumping tube, black         CNY                                  

9232712  Label, BioRobot 9604            CNY                                  

9236429  Sensor, plate switch, TWI            CNY                                  

9239142  Installation, BR MDx DSP            CNY                                  

9239070  Dilutor, Kloehn Versa3           CNY                                  

9232568  Power Cord, USA         CNY                                  

9238970  Target Prep, Premium Agreement  The Premium Cover Agreement TargetPrep provides full service coverage for the BioRobot TargetPrep. Costs for service visits are fully covered including travel, labor, ａnd parts. The agreement also includes one annual preventive maintenance visit, with costs for labor ａnd parts covered. With a Full Agreement, there is a priority response time of 24 hours (1 working day).     CNY                                  

9238791  Angle rotor for 8 Falcon tubes 50 ml           CNY                                  

9232328  Cable, power PCON98 board, BR8000        CNY                                  

9232188  T piece, pressure relief valve, dispense        CNY                            【心肌細(xì)胞總DNA】  

9238906  Upper casing, BS96              CNY                                  

9231863  Elution adapter, auto vac, BR8000        CNY                                  

9238681  Service manual Zent3, german             CNY                                  

9238680  Grease, load-bearing bolts     This is the grease for the Sigma centrifuge. Former cat no 2000770. CNY                                  

9231466  Spacer, DT-Adapter, BR8             CNY                                  

9014582  QIAVac Multiwell Base          CNY                                  

9022195  Reagent Chamber Lid, PyroQ24           CNY                                  

9022022  Sensor, Z-axis, with cable, QXA          CNY                                  

9021653  Kit, stickers, eluate drawer, Qsym        CNY                                  

9021623  Gateway 2, CC-Link, DCU           CNY                                  

9021289  Yellow grease for PM, EZ1           CNY                                  

9240671  Reservoir, 500ml w/cap, AP96/196              CNY                                  

QY-x518  SK-BR-3人乳腺癌細(xì)胞                                   

QY-x519  SK-MEL-1人皮膚黑色素瘤細(xì)胞                                    【心肌細(xì)胞總DNA】

QY-x520  SK-N-SH人神經(jīng)母細(xì)胞瘤                                     

QY-x521  SK-OV-3人卵巢腺瘤細(xì)胞                                      

QY-x522  SMMC-7721人肝癌細(xì)胞                                       

QY-x523  SW13人腎上腺皮質(zhì)瘤                                   

QY-x524  SW480人結(jié)直腸癌                                        

QY-x525  T84人結(jié)腸癌細(xì)胞                                   

QY-x526  THP1人單核細(xì)胞型淋巴瘤                                    

QY-x527  U-2 OS人骨肉瘤細(xì)胞                                     

QY-x528  U251人神經(jīng)膠質(zhì)細(xì)胞瘤                                        

QY-x529  U-937人淋巴瘤細(xì)胞                                       

QY-x530  3T3 swissswiss鼠胚胎成纖維細(xì)胞                                       

QY-x531  3T3L1小鼠胚胎成纖維細(xì)胞(前脂肪)                                   

QY-x532  3T6swiss小鼠胚胎成纖維細(xì)胞                                     

QY-x533  BA/F3小鼠原B細(xì)胞                                      

QY-x534  BHK-21金黃地鼠腎                                        

QY-x535  BS-C-1非注洲綠猴腎                                      【心肌細(xì)胞總DNA】

QY-x536  C2C12小鼠成肌細(xì)胞                                      

QY-x537  C3H 10T1/2 2A6小鼠成纖維細(xì)胞                                         

QY-x538  CHOdhfr二氫葉酸缺陷型中國倉鼠卵巢細(xì)胞                                      

QY-x539  CHO-K1中國倉鼠卵巢細(xì)胞                                   

QY-x540  COS-1非洲綠腎                                      

QY-x541  COS-7非洲綠猴腎細(xì)胞                                         

QY-x542  CV-1猴腎細(xì)胞                                        

QY-x543  IAR20小鼠肝細(xì)胞                                   

QY-x544  IEC-6大鼠小腸隱窩上皮細(xì)胞                                       

QY-x545  LL-PK1,豬腎細(xì)胞                                    

QY-x546  MC3T3-E1小鼠胚胎成骨細(xì)胞                                      

QY-x547  MDCK狗腎細(xì)胞                                     

QY-x548  MEF小鼠胚胎成纖維細(xì)胞