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Answers to many of the frequently asked questions you might have about ADME/Tox products, including tips and tricks for success.

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1.   Where do you supply fresh human hepatocytes? Top
We supply customers throughout North America. At this time, we do not supply fresh human hepatocytes to researchers in countries outside these regions.

2.   Where do you get your liver tissue?
Top
Life Technologies sources liver tissue from a network of hospitals that perform surgical resections on living donors, as well as from deceased donors whose organs were rejected for transplant.  Sourcing for North American customers occurs in North America.

3.   What are the quality specifications on your fresh human hepatocytes?
Top
Our product release specifications include ≥80% viability and ≥80% monolayer confluency (for plated cells).

4.   How often are fresh human hepatocytes available?

Typically, we have access to one or more human liver tissues per week, ensuring a consistent supply.

5.   Are there guidelines for the procurement of human liver tissue? Top

Yes, we comply with country-specific legal and ethical standards for procurement of human liver tissue, including the global ICH Guidelines, and the US’s HIPAA, Uniform Anatomical Gift Act, National Organ Transplant Act, and hospital internal review board (IRB) approval processes.

6.   What is a service level agreement (SLA) and do I need one? Top

 If you are working in a laboratory in the European Union ONLY. An SLA is designed to comply with the UK’s Human Tissue Authority (HTA) regulations and outlines the scope of use for human biologicals.

7.   What is a Rapid Alert? Top

Rapid Alerts notify you of the impending availability of fresh primary human hepatocytes.  Upon receiving a Rapid Alert email, you decide if you need hepatocytes for an upcoming study and can contact us immediately to place an order. Sign up to receive our Rapid Alert.

Each Rapid Alert contains:
- a summary of the donor’s demographics, such as gender, age, race, BMI, and history of alcohol, drugs, or medications
- the plate and suspension formats available for order
- the date when the hepatocytes will be shipped to you
- a phone number and email to place your order or request additional information

8.   How do I sign up for Rapid Alerts? Top
Fill out a Rapid Alert online submission form.

9.   How do I place an order for freshly isolated hepatocytes?
Top
Once you receive a Rapid Alert and determine you want to place an order, reply to the message or call the number listed. Orders are placed prior to the isolation.
Prior to ordering, you must have:
- an approved account with Invitrogen
- a valid purchase order - In North America, purchase orders are made out to Invitrogen. Credit cards are accepted for North American customers.

10.   Can I call Life Technologies Customer Services for fresh hepatocytes? Top
No, the customer service team that helps you order other Life Technologies products does not take orders for fresh hepatocytes. You must respond to the email or phone number that is listed on the Rapid Alert.

11.   If I respond to a Rapid Alert, does that guarantee I will be able to receive fresh human hepatocytes? Top
No, there is no guarantee you will receive hepatocytes. You will not receive fresh human hepatocytes if:
- isolation results do not meet our release criteria
- customer demand exceeds supply

12.   How quickly do I need to respond to a Rapid Alert? Top
Almost all of our orders are taken within four to six hours of a Rapid Alert. To facilitate your order, we recommend a pre-approved blanket purchase order.

13.   Can you notify me of fresh hepatocyte availability through my Twitter account? Top
We are not sending broad-based Twitter alerts at this time. Please discuss any special notification requirements with one of our Hepatic Product Specialists.

14.   How will the fresh hepatocytes be shipped to me? Top
Our hepatocytes are shipped FOB Origin from Durham, NC (North America orders). Fresh suspension cells are sent in cold preservation solution via courier/commercial shippers and delivered as soon as possible. Plated hepatocytes are shipped via courier/commercial shippers one to two days post-plating. This is to ensure good cell health and stabilized CYP450 levels. Courier shipping costs are $300 and commercial shipping costs range between $35 and $100, depending on your region. A standard $92.50 handling fee is added to each order.

15.   How should I handle the fresh hepatocytes once I receive them? Top
Upon receipt, fresh hepatocytes in suspension should be centrifuged at 4ºC immediately, following our instructions. Plated hepatocytes should be quickly removed from the box and the shipping medium replaced with fresh culture medium. The preservation medium used during shipment is meant to be kept cold; if the cells warm past 8ºC, the shipping medium begins to have a toxic effect on the hepatocytes. We recommend that you let your plated cells acclimate overnight before beginning your experiment. Check out our protocol on how to handle fresh hepatocytes in suspension.

16.   Do you sell medium to use with my fresh hepatocytes? Top
Yes, you can order Williams Medium E and supplements for plating and maintaining primary hepatocytes in short-term culture from Invitrogen. Medium can also be ordered and shipped along with your hepatocytes.

17.   Why purchase my fresh hepatocytes with overlay? Top
A more three-dimensional, liver-like framework can be modeled in vitro using a Geltrex overlay, which can extend the life of a healthy hepatocyte culture for more chronic applications such as induction or hepatobiliary transport. A Geltrex overlay is typically recommended for time-points more than 48 hours after isolation. A Geltrex overlay would not be recommended for use before transfecting hepatocytes, or for use in protein secretion studies.

1.   What are the shipping and storage conditions for cryopreserved hepatocytes? Top
Cryopreserved hepatocytes are shipped in the vapor phase of liquid nitrogen contained in a non-hazardous container called a dry LN2 vapor shipper or dewar.  The internal temperature of the dewar is maintained between -140°C and -160°C, generally for up to 5-7 days if unopened.  Once hepatocytes are received and transferred to a long term LN2 dewar  in the laboratory, dewars are returned to the address listed on the pre-paid return shipping label for reuse.

Upon receipt of a shipment of cryopreserved hepatocytes, carefully and quickly transfer the vials to the vapor phase of liquid nitrogen and keep at -135°C or below until use.   Any increase in the temperature of the cryovials before an experiment threatens the viability, functionality, and activity of the hepatocytes.

2.   Do cryopreserved hepatocytes have an expiration date? Top
If properly stored cryopreserved hepatocytes can be stored in the vapor phase of liquid nitrogen (-135°C or below) for several years.  This makes them ideal for a series of experiments conducted over many months.

3.   After thawing, how long can hepatocytes be kept in culture? Top
Unlike immortalized cell lines, hepatocytes are primary cells that cannot be cultured indefinitely.  The use of thawed suspension hepatocytes should be limited to short-term experiments with a maximum of 4-6 hour incubations.  Plateable hepatocytes, which attach to collagen-coated plasticware in culture media, are generally metabolically active for 5-7 days.

4.   Who should I contact if I am looking for hepatocytes with particular characteristics? Top
Our hepatocytes are prequalified and sold according to application, such as induction, short-term metabolism, and transporter uptake.  If you require cells with particular P450 values or donor specifications, contact our Hepatic Biology Tech Support Team or (866) 952-3559 to help with specific lot recommendations.

5.   Who can I call if I have technical questions? Top
Invitrogen has a dedicated team to answer questions regarding hepatocytes. You can send a question by email or contact us by phone:
US toll-free:  +1 866 952 3559
US toll:  +1 919 237 4679