EmbryoMax^® Primary Mouse Embryonic Fibroblasts

Mouse embryonic fibroblasts (MEFs) serve as a feeder layer for both mouse and human embryonic stem cells (ES cells) and induced pluripotent stem cells (iPSCs).

This page covers the indirect co-culture of embryonic stem cells with embryonic fibroblasts.

Stem Cell protocols for cryopreservation, thawing of cryopreserved stem cells and media preparation.

Millipore’s new STEMCCA lentivirus reprogramming kits make it easier than ever to generate induced pluripotent stem (iPS) cells. Unlike traditional iPS generation which requires simultaneous co-infection by four separate expression vectors, the STEMCCA kits use a single polycistronic lentiviral vector to improve efficiency and reduce the number of viral integrations. The STEMCCA vector is comprised of the transcription factors Oct-4, Klf4, SOX-2, and c-Myc (OKSM), separated by the self-cleaving 2A peptide and IRES sequences 1,2. It is also available with flanking LoxP sites incorporated for Cre-mediated excision of the exogenous reprogramming transgenes. STEMCCA Vector Advantages: (1) Efficient: uses a single vector with four transcription factors rather than co-transducing four separate expression vectors (2) Minimizes viral integrations: single vector reduces the risks of insertional mutagenesis and viral reactivation and (3) Excisable: Cre/LoxP-regulated version enables removal of reprogramming transgenes.

"As transgenic mouse core laboratories know well, mouse models are a powerful means of determining the function of mammalian genes in the context of whole organisms—especially since over 95% of the mouse genome is similar to the human genome. However, you may not know that: 1. Laboratory mouse strains are descended from ancestors in modern-day Pakistan. 2. The first inbred strain of lab mouse was created in 1909. It was named DBA for its coat color: dilute brown non-agouti. 3. There is a mutant mouse actually named“Bad Hair Day” (Bhrd). Someone get this little guy a stylist! 4. Male mice, when treated with anticancer drugs, have been shown to pass on DNA instability to their offspring. 5. The founder of Jackson Laboratories, C. C. Little, once had lunch with Walt Disney to request that an animated film be created linking Mickey Mouse and the laboratory mouse. The request was denied."

The cell culture protocols described in this instructional manual include the in vitro culture of murine ES cells using EmbryoMax products along with ESGRO® mLIF medium supplement, as well as feeder-free and serum-free ES cell culture using the ESGRO Complete™ line of products. Also included in this instructional manual are: methods for serum-free neuronal differentiation of mouse ES cells, iPS cell generation using STEMCCA™ lentivirus kits, cre-recombinase mediated excision of STEMCCA™ genes from iPS cells, derivation and rescue of new and existing ES cell lines using RESGRO™ Culture Medium and ESGRO Complete™ system.