Gibco™ HEP10, Pooled Human Cryopreserved Hepatocytes

Catalog No.	Quantity
50-591-338	1.5 mL

Catalog No. 50-591-338 Supplier Gibco™ Supplier No. HMCS10

Description

HEP10, Pooled Human Cryopreserved Hepatocytes

Get the convenience, affordability, and power of a pooled population of hepatocytes in a single vial. HEP10™ Pooled Human Hepatocytes are a representative population of 10 different donor hepatocytes that have been isolated using our novel patent-pending CellStream Isolation Technology™, helping to deliver consistent, predictable results in your drug metabolism studies and other biological assays.

Pure, highly viable hepatocytes isolated using our CellStream Isolation Technology™

10 donor hepatocytes, 5 male and 5 female in each vial

Verified enzymatic and pathway activity

CellStream™ Isolation Technology
HEP10™ Pooled Human Hepatocytes are made using our novel patent-pending CellStream Isolation Technology™. This process eliminates the disadvantages of density gradient fractionation prior to cryopreservation of the pool. Instead, we gently isolate and purify the pre-screened, high-viability, and metabolically-active hepatocytes from other cells and debris. The result is a remarkably pure pooled hepatocyte preparation with high viability for use in drug metabolism studies and other biological assays.

10 Donors in One Vial
Each vial of HEP10™ Pooled Human Hepatocytes contains hepatocytes from ten individual donors, 5 female and 5 male donors. Custom HEP10 lots can also be prepared from our broad array of high-quality, single-donor cells. Large lots are also available for extensive studies.

Verified Activity
Each pool is characterized for CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, CYP3A4⁄5, FMO activity, and NTCP, OATP1B3, and OATP transporter pathways.

Ready for Your Drug Metabolism Experiments
HEP10™ Pooled Human Hepatocytes are useful in a wide variety of applications, including intrinsic clearance, transport, toxicology, biochemistry and cell biology studies.,

Note:
These products require shipment in LN₂ vapor dewars therefore additional fees may apply.

Specifications

• Cell Passage Number: Passage 0
• Content And Storage: Store in a cryostorage dewar or -135°C freezer.
• Cell Type: Hepatocytes
• Culture Type: Suspension Cell Culture
• Form: Cryopreserved
• Gender: Unisex
• Species: Human
• Donor Source: Pooled
• Format: Tube
• No. of Cells: 4-8 x 10⁶
• Product Line: Gibco
• Product Type: Hepatocytes
• Quantity: 1.5 mL
• Tested For: Phase I and II Enzyme Activity, Transporter Suspension Uptake, Viability ≥75%

Frequently Asked Questions (FAQs)

Who can I call if I have technical questions regarding cryopreserved hepatocytes?

Thermo Fisher Scientific has a dedicated team to answer questions regarding hepatocytes. You can send a question by email (hepaticproducts@thermofisher.com) or contact us by phone: US toll-free: (866) 952 3559

Who should I contact if I am looking for hepatocytes with particular characteristics?

Our hepatocytes are prequalified and sold according to application, such as induction, short-term metabolism, and transporter uptake. If you require hepatocytes with particular P450 values or donor specifications, contact our Hepatic Biology Tech Support Team (hepaticproducts@thermofisher.com.com) or (866) 952-3559 to help with specific lot recommendations.

Who can I call if I have technical questions about hepatocyte cell culture?

Thermo Fisher Scientific has a dedicated team to answer questions regarding hepatocyte cell culture. You can send a question by email (hepaticproducts@thermofisher.com) or contact us by phone: US toll-free: (866) 952-3559

Are there guidelines for the procurement of human liver tissue?

Thermofisher Scientific works with a variety of human tissue sources, including Tissue and Organ Procurement Organizations, qualified Research Tissue Organizations and prominent academic and medical centers through collaborations that follow rigorous regulations, certifications and/or accreditations (“Source Facilities”). Tissues obtained through source facilities are consistent with the applicable legal and ethical practices of the United States.

Source Facilities are required by federal law to maintain accurate records and donor confidentiality as well as adhere to certain applicable laws governing tissue used for research globally. These include, but are not limited to:

- 45 CFR Part 46, subparts A, B, C, D where applicable (US federal guidance on the use of human subjects in research)
- Health Insurance Portability and Accountability Act (HIPAA) (addressing issues of confidentiality of personal information)
- National Organ Transplant Act (42 CFR Part 482)
- The Uniform Anatomical Gift Act (UAGA) of 1968, revised 1984

Where do you get your human liver tissue?

The human liver cells are derived from tissue obtained from accredited institutions. Consent is obtained by these institutions from the donor or the donor’s legal next of kin, for the use of the tissue and its derivatives for research purpose.

What should I plate hepatocyte cells on?

Hepatocytes need to be plated on collagen I-coated culture vessels.

I am looking for hepatocytes with particular characteristics. Who should I contact?

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, please contact our Hepatic Biology Tech Support Team at hepaticproducts@invitrogen.com to help with specific lot recommendations.

What is a service level agreement (SLA) and do I need one when working with human tissue?

A service level agreement (SLA) is required ONLY if you are working in a laboratory in the European Union. An SLA is designed to comply with the UK's Human Tissue Authority (HTA) regulations and outlines the scope of use for human biologicals.

Are there guidelines for the procurement of human liver tissue for cryopreserved human hepatocytes?

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.

Where do you get liver tissue for cryopreserved human hepatocytes?

We source liver tissue from a network of hospitals in North America that perform surgical resections on living donors, as well as from deceased donors whose organs were rejected for transplant.

With my hepatocytes, I'm seeing rounding up of the cells, cellular debris, and/or holes in the monolayer, indicating dying cells. What should I do?

This could be due to the toxicity of the test compound. Here are other potential causes and recommendations:

-Sub-optimal culture medium: Use Williams Medium E with Plating and Incubation Supplement Packs; refer to our plating protocol
-Hepatocyte lot not characterized as plateable: Check lot specifications to ensure it is qualified for plating
-Cells were cultured for too long: In general, plateable cryopreserved hepatocytes should not be cultured for more than five days

I'm getting unexpected induction results with my hepatocytes. What could be causing this?

First of all, we recommend comparing results to those reported on our lot-specific characterization specification sheet (human cells) and also referring to our enzyme induction protocol. Here are other potential causes and recommendations:

-Sub-optimal monolayer confluency: Please see our recommendations for 'I have a sub-optimal monolayer confluency for my hepatocytes. What should I do?'
-Poor monolayer integrity: Please see our recommendations for 'With my hepatocytes, I’m seeing rounding up of the cells, cellular debris, and/or holes in the monolayer, indicating dying cells. What should I do?'
-Inappropriate positive control: Check positive control to ensure suitability
-Incorrect concentration of positive control: Use the correct concentration of positive control

I'm seeing sub-optimal bile canlicular formation with my hepatocytes. What should I do?

Please see the following causes and recommendations:

-Hepatocyte lot not transporter-qualified: Check lot specifications to ensure it is transporter-qualified
-Sub-optimal culture medium: Use Williams Medium E with Plating and Incubation Supplement Packs; refer to our plating protocol
-Not enough time for bile canaliculi to form: In general, at least 4-5 days in culture is required for bile canalicular network formation

I'm getting a loss of membrane integrity or cuboidal cell shape with my hepatocytes. What should I do?

Please see the following causes and recommendations:

-Hepatocyte lot not characterized as plateable: Check lot specifications to ensure it is qualified for plating
-Sub-optimal culture medium: Use Williams Medium E with Plating and Incubation Supplement Packs; refer to our plating protocol
-Cells were cultured for too long: In general, plateable cryopreserved hepatocytes should not be cultured for more than five days

I am seeing rounded up cell clumps or debris on top of my hepatocyte monolayer. What should I do?

Please see the following causes and recommendations:

-Seeding density too high: Check lot-specific characterization specification sheet for appropriate seeding density (human cells); observe cells under microscope for appropriate seeding prior to incubation
-Insufficient dispersion of hepatocytes during plating: Disperse cells evenly by moving plate slowly in a figure-eight and back-and-forth pattern in incubator; shake plate and wash cell monolayers prior to applying Geltrex Matrix overlay
-Improper plating volume used for well format : Refer to literature or technical support for suggested plating volumes

I have a sub-optimal monolayer confluency for my hepatocytes. What should I do?

Please see the following causes and recommendations:

-Seeding density too low: Check lot-specific characterization specification sheet for appropriate seeding density (human cells); observe cells under microscope for appropriate seeding prior to incubation
-Insufficient dispersion of hepatocytes during plating: Disperse cells evenly by moving plate slowly in a figure-eight and back and forth pattern in incubator
-Insufficient plating volume used for well format: Refer to literature or technical support for suggested plating volumes
-Low attachment efficiency: Please see our recommendations for: 'I'm getting low attachment efficiency with my hepatocytes. What should I do?'
-Some animal lots are not greater than 80% confluent: Check lot-specific characterization specification sheet for appropriate seeding density. Note: Some animal species create chains or islands of cells rather than being 100% confluent.

I'm getting low attachment efficiency with my hepatocytes. What should I do?

Please see the following causes and recommendations:

-Not enough time for cells to attach: Wait before overlaying with Geltrex Matrix to see if attachment increases; compare cultures to pictures on the lot-specific characterization specification sheet (human cells)
-Poor-quality substratum: Use Gibco Collagen I-Coated Plates
-Hepatocyte lot not characterized as plateable: Check lot specifications to ensure it is qualified for plating; review thawing, plating, and counting protocols

I'm getting an unexpected low hepatocyte cell yield. What could be the cause of this?

Please see the following causes and recommendations:

-Improper thawing technique: Review thawing, plating and counting protocols; thaw cells <2 mins at 37 degrees C
-Sub-optimal thawing medium: Use HTM Medium during thawing to remove cryoprotectant
-Incorrect centrifugation speed: Check thawing protocol for proper centrifugation speed and time (varies by species; human is 100 x g for 10 min at RT)
-Rough handling of hepatocytes during counting: Mix slowly, use wide-bore pipette tips; ensure a homogenous cell mixture prior to counting
-Improper counting technique: Count cells on 2 of the 4 grid lines; do not let cells sit in trypan blue mixture for more than 1 min prior to loading

I'm getting low post-thaw cell viability with my hepatocytes. What should I do?

Please see the following causes and recommendations:

-Improper thawing technigue: Review thawing, plating and counting protocols; thaw cells less than 2 mins at 37 degrees C
-Sub-optimal thawing medium: Use HTM Medium during thawing to remove cryoprotectant
-Rough handling of hepatocytes during counting: Mix slowly, use wide-bore pipette tips; ensure a homogenous cell mixture prior to counting
-Improper counting technique: Count cells on 2 of the 4 grid lines; do not let cells sit in trypan blue mixture for more than 1 min prior to loading
-Cells left out too long: Plate cells immediately after counting

After thawing, how long can hepatocytes be kept in culture?

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 anywhere from 2-7 days depending on which assay they are qualified for use with.

Do cryopreserved hepatocytes have an expiration date?

If properly stored, cryopreserved hepatocytes can be maintained in the vapor phase of liquid nitrogen (-135 degrees C or below) for several years. This makes them ideal for a series of experiments conducted over many months.

What are the shipping and storage conditions for cryopreserved hepatocytes?

Cryopreserved hepatocytes are shipped in the vapor phase of liquid nitrogen contained in a non-hazardous container called a dry liquid nitrogen (LN2) vapor shipper or dewar. The internal temperature of the dewar is maintained between -140 degrees C and -160 degrees C, generally for up to 7-10 days if unopened. Once hepatocytes are received and transferred to a long term LN2 dewar in the laboratory, dewers 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 degrees 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.

Safety and Handling

WARNING: Cancer - www.P65Warnings.ca.gov

For Research Use Only. Not for use in diagnostic procedures.