Gibco™ Mouse (CD-1) Cryopreserved Hepatocytes, Male

Catalog No.	Quantity
MSCS10	1 vial, 1.5 mL

Catalog No. MSCS10 Supplier Gibco™ Supplier No. MSCS10

Description

Mouse (CD-1) Cryopreserved Hepatocytes, Male

These cryopreserved male mouse (CD-1) hepatocytes have been characterized for general Phase I and Phase II drug metabolizing enzyme activities.

Cells per vial: 4-8 million

Get the Results You Need with GIBCO Hepatocytes
High quality hepatocytes, with high viabilities, in vivo-like enzyme expression levels, and proper cell morphology increase the ability to draw in vitro⁄in vivo correlations and make sound decisions regarding a compound's fate. Experienced technicians and proprietary isolation techniques combined with stringent release specifications ensure you receive the highest quality cryopreserved hepatocytes.

Each lot is tested for:

Phase I and II enzyme activities

Proper morphology

Viabilities generally >75%

For research use only. Not intended for human or animal therapeutic or diagnostic use.

Related Links:

Learn more about our cryopreserved hepatocytes

Learn about our full line of hepatocytes ,

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

Specifications

• Content And Storage: Store in a cryostorage dewar or -135°C freezer.
• Cell Type: Hepatocytes
• Culture Type: Hepatocyte Cell Culture
• Form: Cryopreserved
• Gender: Male
• Species: Mouse
• Donor Source: Pooled
• No. of Cells: 4-8 x 10⁶
• Product Line: Gibco
• Product Type: Hepatocytes
• Quantity: 1 vial, 1.5 mL
• Tested For: Phase I and II Enzyme Activity, Proper Morphology, Viability Generally >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

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.

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