PMEF-H
EmbryoMax® Primary Mouse Embryonic Fibroblasts
PMEF, Hygro Resistant, Strain C57/BL6, Mitomycin C Treated, Passage 3
Synonym(s):
MEF Feeder Cells, hygro PMEFs, hygro MEFs, MEFs, Mouse Feeder Cells
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About This Item
biological source
mouse
manufacturer/tradename
Specialty Media
EmbryoMax®
technique(s)
cell culture | stem cell: suitable
input
sample type: mouse embryonic stem cell(s)
sample type primary embryotic fibroblasts (PMEFs)
sample type induced pluripotent stem cell(s)
shipped in
liquid nitrogen
General description
Plating MEF Feeder Cells
Procedure:
1. Prior to thawing PMEF feeder cells, coat plates/flasks with Gelatin solution.
2. Thaw PMEF vial(s) quickly in a 37 °C water bath and transfer to a 15 mL tube (already containing 10 mL of warm PMEF Feeder Cell Medium). Gently invert the tube to distribute, and centrifuge at 300 xg for 4–5 minutes.
3. Remove supernatant and resuspend the cell pellet in warm PMEF Feeder Cell Medium.
4. Remove the Gelatin solution from plates/flasks, and aliquot the PMEF feeder cell suspension at the densities recommended in Table 4.1 of the mouse ES protocol guide
5. Incubate the PMEF Feeder cells at 37 °C with 5% CO2. Use Figures 4A, B and C in the mouse ES protocol guide as a guide for an estimate of correct PMEF density and
appearance. Gelatinized plates may be used for 12–14 days.
Biochem/physiol Actions
Packaging
Preparation Note
Legal Information
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Articles
Mouse embryonic fibroblasts (MEFs) serve as a feeder layer for both mouse and human embryonic stem cells (ES cells) and induced pluripotent stem cells (iPSCs).
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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.
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