Transport Mode
Liquid Nitrogen Transportation
1、Adsorptive Liquid Nitrogen Transportation ,No free liquid nitrogen ,White vapor emission indicates normal operation.
2、Temperature monitoring devices should closely monitor the temperature inside the tank during transportation. If any abnormalities are detected, you can copy the PDF and Excel data from the temperature recorder. (One end of the temperature monitoring device can be unplugged to reveal a USB connector. Simply insert it into a computer to copy the data. If you encounter any difficulties, you can contact the sales staff and technical support for assistance.)
3、Alloy Combination Lock Designed to ensure no third-party access to the LN₂ tank or cell samples during transportation from dispatch to receipt, thereby safeguarding cell integrity.
4、GPS Tracker Enables real-time tracking of cell transportation routes to prevent loss.
5、Liquid Nitrogen Tank, temperature monitoring device, alloy combination lock, and GPS tracker are returnable components. Please store them properly and avoid damage; failure to comply will result in blacklisting.
Reference Citation
pass over.
Scope of Application
Drug Development: Drug metabolism.
Due to biannual updates of our product instruction manual, the following procedures are for reference only.
The most recent version included with the product shall prevail.
II Reagents and Materials
- Primary porcine hepatocytes（Cat# LV-PPH001）
- Recovery medium（Cat#LV-Rec001）
- Purified medium（If desired, give with the cells）
- Plating medium（Adherent cells are needed.Cat#LV-WEP002）
- Sustaining medium（Adherent cells are needed. Cat#LV-WEM002）
- Collagen-coated culture plate（Adherent cells are needed.
Cat #LV-Coated）
- Sterile centrifuge tube of 15 ml
- Disposable pipette
- Wide-mouth pipet tip (The tip of a normal one is cut off and sterilized)
- Pipette
-Thermostat water bath (Preheat at 37°. Please calibrate the temperature with a thermometer)
- Refrigerated horizontal centrifuge (with horizontal rotor. it is possible to centrifuge 15 ml of centrifuge tubes)
- Biosafety cabinet
- 37 °C/5% CO2 incubator
- 75% alcohol
III Resurgence of Suspension Cells
1. In the biosafety cabinet, add 10 ml of resuscitation medium to a 15 ml centrifuge tube and preheat in thermostat water bath of 37 °C for 20 min. Then transfer it to the biosafety cabinet. Plating medium should be preheated at 37 °C.
2. Quickly transfer the frozen hepatocytes from the refrigerated position to a thermostat water bath of 37℃, and immerse them in as much water as possible at 37 °C. Shake it clockwise and horizontally, but make sure that the cap of the freezing tube is kept above the water.
3. Thaw the freezing tube for about 90-120s, until only a small amount of crushed ice floats in it.
4. Sterilize the freezing tube with 75% alcohol and transfer it to a biosafety cabinet.
5. Aspirate the cells with a wide-mouth pipette tip and add them by dropwise to a centrifuge tube containing the preheated recovery medium. (Note: if there are more cells left on the freezing tube and the pipette tip, aspirate 1 ml of resuscitation medium to clean the them. Then, mix them into the medium of the centrifuge tube). Slightly turn the tube upside down 2-3 times to mix well.
6. The survival rate and total number of hepatocytes can be measured by trypan blue exclusion. Add the drug according to the experimental requirements and measure the metabolism.
If the cell density does not meet the requirements or is worried that the cryopreservation has an impact on the activity of metabolic enzymes, it can be directly centrifuged of 50×g at low speed (centrifugation has a certain effect on cell viability), and centrifuged at room temperature for 5min. Go to the supernatant and resuspend with a special medium for drug metabolism (customer-provided). The survival rate and total number of hepatocytes can be measured by trypan blue exclusion. Add the drug according to the experimental requirements and measure the metabolism.
VI Customer Service
If you find any quality problems with the product, please collect the original data and contact the company's salesmen or technical support at the first time. The company ensure after-sales service. Every laboratory has different conditions, different operating habits, different proficiency, and objective factors in experimental failure. If the operation is not strictly in accordance with the instructions or exceeds the time limit of after-sales, the company does not do after-sales. Please understand and support us.
Validity period and raw data provided:
Resuscitation problems: Within 24 hours of resuscitation, trypan blue staining or PI staining should be provided.
Pollution problems: Within 96 hours of resuscitation, microscope photographs of differences should be provided.
Purity issues: Within one month, immunofluorescence or flow cytometry results should be provided
VII Contact Number
Tel:0755-28284050
Technical Support: 19902901483 (Dr. Zhou)
