pluripotent stem cells - derivation
During the window of mammalian preimplantation development, a variety of early embryonic cells or early stage embryos can be used to derive new lines of pluripotent stem cells (PSCs) from mice or humans such as naïve or primed embryonic stem (ES) cells, embryonic germ (EG) cells, and recently expanded potential stem cells (EPSCs). One conventional approach for establishing new ES cell lines is to use the inner cell masses (ICM) of hatched blastocysts: these ICM outgrowths are cultured following immunosurgery to remove the external trophoblasts and expanded to individual ES cell lines. However, noting varying efficiencies, ES cell lines have also been established from early stage embryos including zygotes through hatching blastocysts, parthenotes, individual blastomeres, and explants or dissociated single cells of epiblast. From even later stage E8.5 mouse embryos, another flavor of pluripotent stem cell termed embryonic germ (EG) cell can be derived. Pluripotent stem cells can be established without animal products using defined, xeno-free media and growth substrates. At NovoHelix we have devised several protocols, which also include chemical cocktails, e.g. CHIR99021 (Chiron; CH) and PD0325901 (PD03; PD), that inhibit differentiation pathways to maximize the derivation efficiency of ES cells. We have derived ES cell lines from inbred strains such as C57BL/6J or BALB/c that have traditionally been refractory to colony formation and germline transmission. We have also derived induced pluripotent stem cells from non-permissive strains such as NOD/SHiLtJ. Please see our cell models section on reprogramming for induced pluripotent stem (iPS) cell services.
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Mouse EG (embryonic germ) cell derivation and culture
CMPS001
Derivation of EG cells from E8.5 mouse embryos under 2i-LIF conditions; includes expression testing for Oct4ΔPE-GFP reporter. NovoHelix does not guarantee germline transmission, but many of our EG lines have been injected into C57BL/6 blastocyst to produce high-contribution chimeras.
Derivation of mouse embryonic stem cell line
CMPS002
Re-derive a ES cell line from mouse blastocysts, including ESC karyotype and sex determination. All mESC lines are derived under 2i or 3i media. We provide service of mouse ES cell lines derivation from the strain of choice or background of your mouse line.
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