COMPLETE ES CELL MEDIUM W/ 15% FBS AND LIF

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

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.

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.