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Gibco™ DR4 Mouse Embryonic Fibroblasts, irradiated

Catalog No. A34966
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Quantity:
2 x 106 cells
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Catalog No. Quantity
A34966 2 x 106 cells
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Description

Irradiated Gibco DR4 Mouse Embryonic Fibroblasts (MEFs) are multidrug-resistant feeder cells that support the culture of healthy undifferentiated human and mouse embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs).

Manufactured by MTI-GlobalStem and equivalent to MTI-GlobalStem Cat. No. GSC-6204G, these MEF cells are isolated from DR4 x CF1 hybrid mice, mitotically arrested by irradiation, and tested to help ensure safety and performance. For other variants of drug-resistant MEFs, see CF6-Neo Mouse Embryonic Fibroblasts, Irradiated or CF6-Neo Mouse Embryonic Fibroblasts, MitC-Treated. You may also choose from our selection of non-drug-resistant MEFs to suit your application.

• Help save precious time with pre-inactivated, ready-to-use MEFs
• Culture ESCs and iPSCs with confidence using meticulously tested feeder cells
• Perform drug selection with MEFs that are resistant to neomycin/geneticin, puromycin, hygromycin, and 6-thioguanine

Help save time with ready-to-use MEFs
Irradiated DR4 MEFs help save time and trouble dealing with an animal facility, performing tedious, time-consuming dissections, as well as expanding and testing cells day after day, week after week. Focus on your research with these pre-inactivated, ready-to-use feeders.

Culture with meticulously tested feeder cells
Each lot of DR4 MEFs is tested for bacteria, fungi, mycoplasma, and various mouse pathogens to minimize the risk of contamination. Each lot is also tested for post-thaw viability (typically greater than 95%) and ability to support mouse and human ESCs to help ensure robust and consistent performance. These MEFs have successfully been used with ESC/iPSC lines from other species, but should be tested for compatibility if used for such an application.

Perform drug selection with multidrug-resistant cells
DR4 MEFs express transgenes that make them resistant to four different drugs: geneticin (G418), puromycin, hygromycin, and 6-thioguanine. Each lot is tested for tolerance to various concentrations of these drugs and the threshold range for survival is reported in the certificate of analysis to aid in designing experiments. Use these feeder cells to support undifferentiated ESC or iPSC culture while performing drug selection.

Specifications

Content And Storage Cryopreserved mouse embryonic fibroblasts

Store in liquid nitrogen.
Age Embryonic
Cell Type Feeder Cells (Mouse Embryonic Fibroblast)
Form Cryopreserved
Species Mouse
Donor Source Pooled
No. of Cells 2 x 106
Quantity 2 x 106 cells
Shipping Condition Shipped on dry ice

Frequently Asked Questions (FAQs)

Is it better to use irradiated mouse embryonic fibroblasts (MEFs) or Mitomycin C-treated MEFs to grow embryonic stem cells?

Either method will work in arresting cell division. However, the irradiation process will ensure that cell division will cease regardless of cell aggregation. Cell clumping can potentially not inactivate all cells when using mitomycin C, as cells within clumps may not be exposed to the mitomycin C. Irradiated cells are preferred by those who have concerns about chemical treatment. Mitomycin C-treated cells are preferred by those who have concerns about DNA damage from irradiation.
What are the differences between CF1, CF6, and DR4 mouse embryonic fibroblasts?

CF1 mouse embryonic fibroblasts do not have drug resistance. CF6 mouse embryonic fibroblasts are resistant to Neomycin/geneticin (G418). DR 4 mouse embryonic fibroblasts are resistant to geneticin (G418), puromycin, hygromycin, and 6-thioguanine.

What is the recommended seeding density for mouse embryonic fibroblasts? Why is the seeding density critical?

We recommend seeding these cells at densities ranging from 2 x 10E4 to 5.5 x 10E4 cells/cm2. A good starting point is 3 x 10E4 cells/cm2. If the feeder cells are too sparse, they may not maintain the pluripotent cells without differentiation, and the pluripotent cells may not attach well. If the feeder cells are too dense, the feeder layer may detach from the plate, and the culture will be lost.
When should mouse embryonic fibroblasts be plated and how long can these cells be used?

These cells should be plated 24 hours prior to plating the ESCs or iPSCs and should be used for only 7-10 days.
Does the culture vessel need to be coated prior to seeding mouse embryonic fibroblasts?

Yes, the culture vessel needs to be coated with Attachment Factor protein (Cat. No. S006100) at 37 degrees C for 30 mins or at room temperature for 2 hours. The coated vessels can be used immediately or stored at room temperature for up to 24 hours.

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