Two stages of mouse pluripotency have been stabilized to date through in vitro culture. ‘Naive’ represents the earliest stage, which can be enriched for a purer population, termed ‘ground state’. Naive most closely approximates embryonic epiblast in the preimplantation blastocyst. The primed state most closely approximates in vivo immediate postimplantation epiblast. There are clear differences between these stages in culture, expression, epigenetics (including appropriate X-inactivation) and developmental competence. The fact that naive mESCs can generate whole animals through tetraploid complementation while primed mESCs cannot, is a particularly compelling reason for heightened expectations from naive hESCs.
ESGRO-2i medium is a defined, LIF containing medium provided with a selective GSK3β and Mek 1/2 inhibitors to enhance viability of mouse embryonic stem (ES) and induced pluripotent stem (iPS) cells and increased maintenance of pluripotency in the absence of serum and feeder cells. This medium is intended for cell expansion and growth at clonal density in serum-free conditions and supports a naive/ground pluripotent state.
Suitable for ES and iPS cell derivation, passaging and maintenance. Media can be used on feeder dependent and independent pluirpotent stem cell lines.
Important passaging information
Use of one of the below listed cell dissociation solutions is imperative while passaging ES cells. Standard trypsin protocols used to passage ES cells grown in serum containing media typically result in over trypsinization causing the cells to lift off the plates and form abnormal ball-like structures.
Recommended products include:
ESGRO Complete™ Accutase (Cat # SF006)
ESGRO Complete Trypsin Solution (proprietary) (Cat # SF007)
ESGRO Complete Enzyme-Free Dissociation Solution (proprietary) (Cat # SF009)
1. ESGRO-2i Basal Medium with LIF: (Catalogue No. CS204463) One (1) 100 mL bottle containing basal medium supplemented with LIF, pH 7.1-7.3.
2. GSK3β Inhibitor Supplement: (Catalogue No. SF012-050) One (1) 50 μL vial containing selective GSK3β inhibitor supplement.
3. Mek1/2 Inhibitor Supplement: (Catalogue No. CS204465) One (1) 10 μL vial containing selective Mek 1/2 inhibitor supplement.
Storage and Stability
• Upon receipt, store basal medium and inhibitor supplements in the dark at -20°C until use. Refer to lot expiration date on labels.
• After basal medium and inhibitor supplements are combined, store the complete ESGRO-2i medium in the dark at 4°C for up to 10 days. Do not refreeze.
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