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A propos de cet article
biological source
mouse
Quality Segment
manufacturer/tradename
Specialty Media, EmbryoMax®
technique(s)
cell culture | stem cell: suitable
input
sample type: human embryonic stem cell(s)
sample type: mouse embryonic stem cell(s)
sample type primary embryotic fibroblasts (PMEFs)
sample type induced pluripotent stem cell(s)
shipped in
liquid nitrogen
storage temp.
-140 to -196°C
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
Physical form
Preparation Note
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Cet article | |||
|---|---|---|---|
| technique(s) cell culture | stem cell: suitable | technique(s) cell culture | stem cell: suitable | technique(s) cell culture | stem cell: suitable | technique(s) cell culture | stem cell: suitable |
| biological source mouse | biological source mouse | biological source mouse | biological source mouse |
| Quality Level 100 | Quality Level 100 | Quality Level 100 | Quality Level 100 |
| shipped in liquid nitrogen | shipped in liquid nitrogen | shipped in - | shipped in liquid nitrogen |
| storage temp. -140 to -196°C | storage temp. -140 to -196°C | storage temp. - | storage temp. -140 to -196°C |
| manufacturer/tradename Specialty Media, EmbryoMax® | manufacturer/tradename Specialty Media, EmbryoMax® | manufacturer/tradename Specialty Media, EmbryoMax® | manufacturer/tradename Specialty Media, EmbryoMax® |
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Classe de stockage
12 - Non Combustible Liquids
wgk
WGK 2
flash_point_f
does not flash
flash_point_c
does not flash
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Protocoles
Stem Cell protocols for cryopreservation, thawing of cryopreserved stem cells and media preparation.
This page covers the indirect co-culture of embryonic stem cells with embryonic fibroblasts.
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).
Contenu apparenté
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.
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.
Numéro d'article de commerce international
| Référence | GTIN |
|---|---|
| PMEF-CFL | 04053252552045 |
