Can KnockOut™ SR be used to culture mouse ESCs on feeder cells?

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Yes, KnockOut™ SR was designed to culture mouse ES cells in undifferentiated conditions.

Answer Id: E4974

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How do I characterize human embryonic stem ( ES) cells?

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Human ES cells are generally characterized by their typical morphology (they grow as tightly packed clusters of small cells with high ratio of nucleus to cytoplasm); surface marker expression; RT-PCR detection of stem cell-specific gene expression (such as Oct3/4, Sox2, and Nanog); alkaline phosphatase staining, and telomerase activity assay. The most commonly used ES specific surface markers include stage-specific embryonic antigens SSEA-3 and SSEA-4 for human ES cells. Other ES-specific surface antigens also include TRA-1-60 and TRA-1-81. (Science 282:1145 (1998).

Answer Id: E12179

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Can KnockOut™ SR be used to culture mouse ESCs in feeder-free conditions?

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Yes, mouse embryonic stem cells can be cultured using KnockOut™ SR under feeder-free conditions. Typically, cells are plated at a higher seeding density, in the presence of LIF, on a 0.1% gelatin layer in place of feeder cells.

Answer Id: E4977

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How are human embryonic stem (ES) cells derived?

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Human ES cells are derived from human blastocyst inner cell masses, isolated by immunosurgery with rabbit antiserum to BeWO cells (a human trophoblast cell line) (Science 282:1145 (1998)).

Answer Id: E12177

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How do I grow human embryonic stem (ES) cells in vitro?

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Human ES cells are typically maintained on top of irradiated mouse embryonic fibroblast cells (MEF) with high-glucose DMEM supplemented with 20% FBS (or 20% of Gibco™ KnockOut™ Serum Replacement), 0.1 mM beta-mercaptoethanol, 1% of nonessential amino acids (Science 282:1145 (1998); Dev Biol 227:271 (2000)).

Human ES cells can also be maintained without feeder cells on Matrigel™ matrix–coated culture vessels in medium conditioned by MEF (Nat Med 10:55 (2004)). Once the ES cells are established on the MEF or MEF-conditioned medium, these cells may be maintained under feeder-free conditions using Gibco™ Geltrex™ Matrix-coated culture vessels in Gibco™ StemPro™ SFM supplemented with bFGF and beta-mercaptoethanol or Gibco™ Essential 8™ Medium and vitronectin-coated vessels.

Answer Id: E12178

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What are ES cells?

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Embryonic stem (ES) cells are derived from the early mammalian embryo and are capable of unlimited, undifferentiated proliferation in vitro while maintaining their potential to differentiate into a wide of range of adult tissues including germ cells. The pluripotency of the ES cells is normally demonstrated in vitro by inducing ES cells to differentiate into embryoid bodies and checking lineage-specific markers for differentiated cells in three body layers (endo, meso, and ectoderm), or injecting them into immunodeficient mice and determining the cell types produced in the teratomas.

Answer Id: E12176

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