Anti-Stage-Specific Embryonic Antigen-1 Antibody, clone MC-480

Traditionally, culture and expansion of mouse embryonic stem cells (mESC) is performed using several single-layer T-flasks or Petri dishes. Harvesting and analyzing stem cells from multiple culture vessels is a repetitive and time-intensive process that consumes valuable incubator space. Here, we report a method for expanding mESCs using a new multilayer cell culture flask. The Millicell HY multilayer culture flask makes expansion and harvesting of mESCs faster and easier than traditional culture methods. The method for using the new flasks is linearly scalable per unit surface area. Therefore, no reoptimization of procedure is required – culture conditions and media volume are identical, regardless of whether culture is performed in traditional single-layer devices or in a Millicell HY culture flask. Resulting morphology, cell recovery, viability, and pluripotency of mESCs were not significantly different between multilayer flask culture and single-layer flask culture methods.

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.