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Mouse Embryonic Fibroblast (MEF) Conditioned Media
R&D Systems, Inc. a Bio-Techne Brand
Key Product Details
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Induced Pluripotent Stem Cells Grown in MEF Conditioned Media Express Pluripotent Stem Cell Markers SSEA-4, Oct-3/4, Oct-4A, and E-Cadherin. Induced Pluripotent Stem Cells Grown in MEF Conditioned Media Express Pluripotent Stem Cell Markers SSEA-4. Oct-3/4, Oct-4A, and E-Cadherin.Human iPS2 stem cells were cultured in MEF Conditioned Media (Catalog # AR005) supplemented with 4 ng/mL Recombinant Human FGF basic (Catalog # 233-FBor Catalog # 4114-TC).A. Expression of SSEA-4 was detected using a Mouse Anti-Human/Mouse SSEA-4 Monoclonal Antibody (Catalog # MAB1435) followed by a NorthernLights™ (NL) 493-conjugated Donkey Anti-Mouse Secondary Antibody (Catalog # NL009, green). Expression of Oct-3/4 was detected using a Goat Anti-Human Oct-3/4 Antigen Affinity Purified Polyclonal Antibody (Catalog # AF1759) followed by NL557-conjugated Donkey Anti-Goat IgG (Catalog # NL001, red). The nuclei were counterstained with DAPI.B. Expression of Oct-4A was detected using Mouse Anti-Human Oct-4A Monoclonal Antibody (Catalog # MAB17591) followed by a NorthernLights™ 493-conjugated Donkey Anti-mouse Polyclonal Antibody (Catalog # NL009, green). Expression of E-Cadherin was detected using a Goat Anti-Human E-Cadherin Antigen Affinity Purified Polyclonal Antibody (Catalog # AF648) followed by a NL557-conjugated Donkey Anti-Goat IgG Secondary Antibody (Catalog # NL001, red). The nuclei were counterstained with DAPI. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips. |
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Human Emryonic Fibroblast Conditioned Media Express Pluripotent Markers SSEA-4 and Oct-3/4. Human Emryonic Fibroblast Conditioned Media Express Pluripotent Markers SSEA-4 and Oct-3/4. A.Human embryonic stem cells were cultured in Mouse Embryonic Fibroblast (MEF) Conditioned Media (Catalog # AR005) supplemented with 4 ng/mL Recombinant Human FGF basic (Catalog # 233-FB). SSEA-4 and Oct-3/4 were detected using a Mouse Anti-Human/Mouse SSEA-4 Monoclonal Antibody (Catalog # MAB1435, red) and a Goat Anti-Human Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759, green). The nuclei were counterstained with DAPI (blue).B.Human embryonic stem cells were cultured in MEF Condition Media supplemented with 4 ng/mL Recombinant Human FGF basic. SSEA-4 and Oct-3/4 were detected as described above. The nuclei were counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips. |
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, MEF Conditioned Media can be used to culture human pluripotent stem cells using the following procedure:
- Supplement MEF Conditioned Media with 4 ng/mL FGF basic
- Culture cells in supplemented media
- Replace media daily and passage cells as needed
Reagents Provided
Reagents Supplied in Mouse Embryonic Fibroblast (MEF) Conditioned Media (Catalog # AR005)
- 100 mL MEF Conditioned Media
Other Supplies Required
Reagents
- Recombinant Human FGF basic (Catalog # 233-FB) or tissue culture grade Recombinant Human FGF basic (Catalog # 4114-TC)
- Accutase® (Innovative Cell Technologies)
- Cultrex® Reduced Growth Factor Basement Membrane Extract (BME) (Catalog # 3433-005-01)
- DMEM/F12
Materials
- BG01V human embryonic stem cells or equivalent
- 60 or 100 mm tissue culture plates
- 15 mL centrifuge tubes
- Pipettes and pipette tips
Equipment
- 37 °C and 5% CO2 incubator
- Centrifuge (low speed clinical or equivalent)
- Hemocytometer
- Microscope
Procedure Overview
Preparation of BME-coated Plates
- Thaw Cultrex BME on ice at 2 °C to 8 °C overnight.
- Aliquot thawed Cultrex BME into pre-cooled tubes and store at -20° C.
- Thaw the aliquot on ice at 2 °C to 8 °C.
- Dilute Cultrex BME 1:40 in DMEM/F12. This can be stored at 4 °C for up to 2 weeks.
- Coat the desired number of plates with diluted Cultrex BME (approximately 2.5 mL/60 mm plate) and incubate for 1-2 hours at room temperature.
- Remove the Cultrex BME solution immediately prior to plating the cells.
I. Preparation and plating of BG01V Human Embryonic Stem Cells
Gently transfer thawed BG01V human embryonic stem cells to a 15 mL centrifuge tube containing pre-warmed MEF Conditioned Media.
Centrifuge at 200 x g for 4 minutes.
Resuspend the cell pellet in MEF Conditioned Media supplemented with 4 ng/mL FGF basic.
Add the BG01V human embryonic stem cells to the Cultrex BME- or StemXVivo-Culture Matrix-coated plates.
Culture the cells at 37 °C and 5% CO2 and change the media daily.
II. Passaging BG01V Human Embryonic Stem Cells
Coat plates with Cultrex BME or StemXVivo Culture Matrix as described.
Remove the media from cells and add 1 mL of Accutase solution to each 60 mm plate.
Pipette gently over the plate until all of the cells have detached.
Transfer the cell suspension to a 15 mL centrifuge tube containing < 5 mL of MEF Conditioned Media.
Centrifuge at 200 x g for 4 minutes.
Resuspend the cell pellet in MEF Conditioned Media.
Perform a cell count.
Plate the desired number of cells (approximately 1.0 x 106 cells/60 mm plate) on Culture BME- or StemXVivo Culture Matrix-coated plates in MEF Conditioned Media containing 4 ng/mL FGF basic.
Culture the cells at 37 °C and 5% CO2 and change the media daily.
BG01V cells are licensed from ViaCyte, Inc.
Citations for Mouse Embryonic Fibroblast (MEF) Conditioned Media (6)
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Citations are publications that use Bio-Techne products. Selected citations for Mouse Embryonic Fibroblast (MEF) Conditioned Media include:
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Species: Northern White Rhinoceros
Sample Types: Whole Cells
Applications: BioassayV Zywitza et al. (2022), Na�ve-like pluripotency to pave the way for saving the northern white rhinoceros from extinction Scientific Reports, 12(1):3100.
PMID: 35260583 -
Species: Mouse
Sample Types: Media
JC Chiang et al. (2018), Trisomy silencing by XIST normalizes Down syndrome cell pathogenesis demonstrated for hematopoietic defects in vitro Nat Commun, 9(1):5180.
PMID: 30518921 -
Species: Human
Sample Types: Whole Cells
C Ness et al. (2017), Multicellular tumor spheroids of human uveal melanoma induce genes associated with anoikis resistance, lipogenesis, and SSXs Mol. Vis., 23(0):680-694.
PMID: 29033534 -
Species: Human
Sample Types: Whole Cells
Zhou X et al. (2014), Modulating innate immunity improves hepatitis C virus infection and replication in stem cell-derived hepatocytes. Stem Cell Reports, 3(1):204-14.
PMID: 25068132 -
Species: Mouse
Sample Types: Whole Cells
Cheli Y et al. (2011), Mitf is the key molecular switch between mouse or human melanoma initiating cells and their differentiated progeny. Oncogene, 30(20):2307-18.
PMID: 21278797 -
Species: Human
Sample Types: Whole Cells
Mandal S et al. (2011), Mitochondrial function controls proliferation and early differentiation potential of embryonic stem cells. Stem Cells, 29(3):486-95.
PMID: 21425411
There are no citations that match your criteria.
Customer Reviews for Mouse Embryonic Fibroblast (MEF) Conditioned Media (2)
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Verified Customer | Posted 09/13/2019We used to make are own, however the quality would often vary. This commercial media enables us to standardise the protocol and we have used a number of lots with equal results. We usually use the media for 24 hours before starting a iPSC differentiation in order to 'prime' the cells. I would recommend for those who wish to increase the efficiency of iPSC differentiation.
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Verified Customer | Posted 04/04/2018
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FAQs for Mouse Embryonic Fibroblast (MEF) Conditioned Media
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Have you tried the StemXVivo® Cardiomyocyte Differentiation Kit with other human ES cell lines or human iPS cell lines?
This kit has been tested both in-house and by outside groups in multiple iPS cell lines. This kit is expected to work with most cell lines as long as the starting stem cell population is of high quality and has pluripotency. For optimal performance using this kit, it is recommended that pluripotent stem cells are grown in Mouse Embryonic Fibroblast (MEF) Conditioned Media (Catalog # AR005), however successful differentiation has been observed in cells grown with mTeSR™ medium. We have had difficulties with cells grown in TeSR™-E8 media. An anti-human Cardiac Troponin T antibody is provided in the kit to help determine the efficiency of differentiation.
