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Mouse Embryonic Blasticidin and Zeocin-resistant (MMMbz) Immortalized Feeder Ce

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Mouse Embryonic Blasticidin and Zeocin-resistant (MMMbz) Immortalized Feeder Ce

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Mouse Embryonic Blasticidin and Zeocin-resistant (MMMbz) Immortalized Feeder Cells

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Safety	Biosesafety Level II
Organism	NMR1 Mouse
Source Organ	Embryo
Growth Properties	Adherent
Morphology	Fibroblast-like
Applications	For Research Use Only.
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Description	Supporting cells, such as short-term cultures of fibroblasts, are used to cultivate human embryonic stem (ES) cells, however they are susceptible to senescence after a number of passages. The Mouse Embryonic Blasticidin and Zeocin-resistant (MMMbz) Immortalized Feeder Cells eliminates the problems associated with the use of primary cell lines. This cell line has a doubling time of 16 hours and may be used indefinitely to propagate human embryonic stem cells. It has the ability to facilitate the growth of hES H9 and hES H1. MMMbz cells are resistant to blasticidin and zeocin due to transgene expression allowing for selection of hES cells transfected with genes conferring resistance.
Quality Control	Selective growth with media containing blasticidin and zeocin to select for cells with transgene expression.
Propagation	Grow cells in T25 ECM-coated flasks (G299) with the following conditions. The base medium for this cell line is Prigrow III medium available from ABM (TM003). To make the completed growth medium, add the following components to the base medium: Bovine Calf Serum (available from gibco, SH30072) to final concentration of 10% and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO₂: 5%; Temperature: 37.0°C. To freeze: suspend cells in PriGrow III (TM003) with 10% bovine calf serum and 10% sterile DMSO. To support the growth of hES cells: irradiate MMMbz cells with 6000 rads and seed at a density of 1.3x10⁵/cm² in propagation medium. The next day add hES in Knockout DMEM (Invitrogen), supplemented with 20% Knockout Serum Replacement (Invitrogen), 1mM L-glutamine (G275), 1% MEM non-essential amino acids (Invitrogen), 0.1mM 2-mercaptoethanol (CH045) and bFGF at 4 ng/ml (Z200015).
Storage	-180°C
Shipping	On Dry Ice
Notes	For Research Use Only, not for therapeutic or diagnostic purposes.