Feeder layers of mouse embryonic fibroblasts (MEF) support stem cell growth and the maintenance of pluripotency by secreting growth factors, cytokines, and nutrients. However, the use of MEFs as a feeder layer can be laborious and MEF contamination is a significant concern. See Details
MEF conditioned media represents an efficient alternative to MEF feeder cells and is one of the mostly widely used feeder-free systems for embryonic stem cell culture. MEF Conditioned Media includes the soluble factors required to support stem cell growth and pluripotency.
Upon receipt, store the conditioned media at < -20 °C in a manual defrost freezer. When ready to use, thaw the solution overnight at 2 °C to 8 °C in the dark. Aliquot and store the unused portions at < -20 °C in a manual defrost freezer. Long-term storage at 2 °C to 8 °C is not recommended. Avoid repeated freeze-thaw cycles, and do not use past the expiration date.
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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. Human iPS2 stem cells were cultured in MEF Conditioned Media (Catalog # AR005) supplemented with 4 ng/mL Recombinant Human FGF basic (Catalog # 233-FB or 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 Embryonic Stem Cells Cultured in Mouse Embryonic 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.
Image courtesy of Dr. Frank Soldner of the National Institutes of Health.
Embryonic stem (ES) cells are pluripotent stem cells derived from the inner cell mass of pre-implantation embryos. Induced pluripotent stem (iPS) cells can be generated by somatic cell reprogramming following the exogenous expression of specific transcription factors (Oct-3/4, KLF4, SOX2, and c-Myc). These cell types are capable of unlimited, undifferentiated proliferation in vitro and still maintain the capacity to differentiate into a wide variety of somatic cells. In this capacity, pluripotent stem cells have widespread clinical potential for the treatments of heart disease, diabetes, spinal cord injury, and a variety of neurodegenerative disorders.
R&D Systems offers a wide range of products to support pluripotent stem cell culture and differentiation. Mouse embryonic fibroblasts may be used to maintain and expand pluripotent stem cells in an undifferentiated state. We also offer defined culture media, which are specifically optimized for use with human or rodent pluripotent stem cells. In addition, R&D Systems offers a variety of products to assess differentiation status and identify specific stem cell types of interest, including panels of marker antibodies, primer pairs, multi-color flow cytometry kits, and specialized verification kits.
Refer to the product datasheet for complete product details.
Reagents Supplied in Mouse Embryonic Fibroblast (MEF) Conditioned Media (Catalog # AR005)
Prepare Either A) Cultrex BME-coated plates or B) StemXVivo Culture Matrix-coated Plates
A. Preparation of BME-coated Plates
B. Preparation of StemXVivo Culture Matrix-coated Plates
I. Preparation and plating of BG01V Human Embryonic Stem Cells
II. Passaging BG01V Human Embryonic Stem Cells
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