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Mouse Embryonic Stem Cell Culture

 Transgenic Mouse Models

Transgenic and gene knockout technologies are powerful tools for studying gene function. A commonly used method for creating transgenic and knockout mice involves the introduction of genetically modified embryonic stem cells into early-stage mouse embryos by either blastocyst injection or aggregation techniques. These methods result in the generation of chimeric offspring; the genetic modification may be transmitted to successive generations if the ES cells contribute to the germline.

MilliporeSigma offers a broad range of tools and technologies to culture mouse embryonic stem cells including the gold standard ESGRO/mLif supplement, ESGRO complete and ESGRO2i serum-free/feeder-free media. We also offer commonly used mouse embryo media and MEF feeder cells for the storage, transfer and expansion of mouse embryos used to create transgeneic mouse models under the EmbryoMAX™ name.

Browse all Mouse Embryo Validated Reagents

Join Researchers at the Cutting Edge of Genome Editing


Generating Mouse Models using Embryonic Stem Cells

Figure 1. General procedure for generating genetically modified mice. Pluripotent ES cells can be purchased or isolated from mouse blastocysts. The ES cell genome can be modified in two ways: 1) add expression of an exogenous gene to the ES cell 2) disrupt the expression of an endogenous gene. In both cases, the targeting vector contains sequences homologous to the target gene as well as sequences changing the target and enabling positive/negative selection. Modified ES cells are microinjected into blastocysts. The new CRISPR/Cas9 genome editing system provides the option of directly microinjecting zygotes with the targeting mRNAs along with the Cas9 mRNA. Blastocysts are injected into the mouse to generate chimeric mice. These are mated with normal mice to generate mice with the desired genetic modification.

 Mouse Embryonic Stem Cell Culture

ESGRO® mLIF Supplement

Maintaining pluripotency for over a decade

For over a decade, stem cell researchers have trusted their cultures with ESGRO® mouse LIF supplement for maintaining the pluripotent state of their mouse ES cell lines. The gold standard for undifferentiated mouse ES cell culture, ESGRO® mLIF features:

  • Consistent inhibition of ES cell differentiation
  • Convenient format, supplied in active units/mL
  • No batch-to-batch variation
  • Flexibility; supports feeder-free and feeder-based cell culture


Alkaline phosphatase staining shows that
ESGRO® supplement inhibits mES cell
differentiation.
 


Ordering Info
 

Product No.
Description
ESG1106
ESGRO® Leukemia Inhibitory Factor (LIF), 1 million units/1 mL
ESG1107 ESGRO® Leukemia Inhibitory Factor (LIF), 10 million units/1 mL
L5158 Leukemia Inhibitory Factor Protein, Recombinant mouse
L5283 Leukemia Inhibitory Factor Protein, Recombinant human
L9545 Leukemia Inhibitory Factor human, animal component free

 Serum-Free and Feeder-Free Media

ESGRO Complete™ PLUS Media
The ESGRO Complete™ system is the first to enable serum-free and feeder-free culture of mouse ES cells. The cornerstone of this system is the ESGRO Complete™ clonal grade medium, which supports the self-renewal of mouse ES cells by providing the basic nutrients normally supplied by serum and feeders in the traditional culturing method. These nutrients include hormones, vitamins, the growth factors mLIF and BMP4, as well a selective GSK3β inhibitor for enhanced mouse ES cell growth and viability at clonal densities in serum-free conditions.

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ESGRO®-2i Medium
Based on the principle of using LIF along with GSK3β and Mek 1/2 small molecule inhibitors to establish and sustain ES cells in a “ground state of pluripotency1-3”, The ESGRO®-2i medium or supplement is designed to maintain “naïve state” or “ground state” culture conditions, enhance viability and ease of cell culturing of ES cells and Increases maintenance of pluripotency without serum and feeder cells

Application Note: Naive Pluripotent Stem Cell Culture in a Novel Inhibitor Based 2i/LIF Containing Serum-Free Stem Cell Media

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Mouse ES cells grown in ESGRO®-2i medium express the pluripotency markers Oct-4, Sox-2 and SSEA1 (left to right).

Browse all Mouse ES Cell Lines

Browse all EmbryoMax Reagents

 MEF Feeder Cells

The EmbryoMax range of PMEF cells provides researchers with a convenient solution for ES cell culture by eliminating the need for time-consuming MEF feeder cell isolation and preparation. Derived from day 13 embryos, these cells are supplied frozen at passage three (2 populations doublings per passage) in five-vial packs. Each vial contains approximately 5-6 x 106 fibroblasts.
 

Product Description
Treatment Passage Qty/Pk Product No.
EmbryoMax® Primary Mouse Embryo
Fibroblasts Strain CF1
Not Treated

Not Treated

Mitomycin-C Treated

Irradiated
P1

P3

P3

P3
1 vial, 1 x 106 cells ea.

5 vials, 5-6 x 106 cells ea.

5 vials, 5-6 x 106 cells ea.

5 vials, 5-6 x 106 cells ea.
PMEF-CFL-P1

PMEF-CFL

PMEF-CF

PMEF-CFX
EmbryoMax® Primary Mouse Embryo
Fibroblasts Neo Resistant
Not Treated

Not Treated

Mitomycin-C Treated

Irradiated
P1

P3

P3

P3
1 vial, 1 x 106 cells ea.

5 vials, 5-6 x 106 cells ea.

5 vials, 5-6 x 106 cells ea.

5 vials, 5-6 x 106 cells ea.
PMEF-NL-P1

PMEF-NL

PMEF-N

PMEF-NX
EmbryoMax® Primary Mouse Embryo
Fibroblasts Hygro Resistant
Not Treated

Mitomycin-C Treated
P3

P3
5 vials, 5-6 x 106 cells ea.

5 vials, 5-6 x 106 cells ea.
PMEF-HL

PMEF-H

 Mouse Genotyping

KOD Xtreme™ Hot Start DNA Polymerase

DNA Amplification from Challenging Crude Samples with Minimal Processing

The KOD Xtreme™ Hot Start DNA Polymerase is your enzyme of choice for the most challenging PCR situations including: DNA amplification from crude samples such as mouse tail tip lysates, high GC content, or repeat sequences (T/A) which can inhibit or bias PCR amplification data.

Features and Benefits:

  • Optimized for PCR success against complex crude samples and with minimal processing
  • Efficiently amplifies up to 90% GC-content templates
  • 10x higher fidelity than Taq blends
  • Amplifies genomic targets up to 24kb
  • Amplifies plasmid/phage targets up to 40kb
  • Eliminate mispriming or primer-dimer formation
  • Convenient ambient temperature setup compatible with automation

Amplify DNA from crude tissue lysate with minimal processing to save time and minimize cost while maintaining quality data.

  The targeted locus genes were amplified with various PCR enzymes using mouse tail lysates prepared by alkaline lysis*. KOD Xtreme™ Hot Start DNA polymerase successfully amplified both targets (Tg and WT).

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