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Differentiation of Embryonic Stem (ES) Cells Using the Hanging Drop Method - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4485

Category: Stem Cell Protocols: Stem Cell Culture Protocols

In vitro differentiation of ES cells occurs when the cells are allowed to aggregate in suspension culture in the absence of mouse embryonic fibroblast (MEF) feeders and leukemia inhibitory factor (LIF). Hanging drops provide a uniform aggregate size, which is then expanded by continued growth in suspension culture. The embryoid bodies are then plated and allowed to differentiate further in culture. - [Read Differentiation of Embryonic Stem (ES) Cells Using the Hanging Drop Method]

Isolation and Freezing of Primary Mouse Embryonic Fibroblasts (MEF) For Feeder Plates - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4482

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the isolation of fibroblasts from mouse embryos. Mouse embryonic fibroblast (MEF) cells are used as a feeder layer for the culture of mouse embryonic stem (ES) cells to help maintain them as pluripotent stem cells. The inhibition of ES-cell differentiation provided by the MEF feeders appears to be due to their production of leukemia inhibitory factor (LIF). - [Read Isolation and Freezing of Primary Mouse Embryonic Fibroblasts (MEF) For Feeder Plates]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the culture of embryonic stem (ES) cells using mitotically inactivating primary mouse embryonic fibroblast (MEF) cells as a feeder layer (preparation described in Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates). The ES culture medium is supplemented with recombinant leukemia inhibitory factor (LIF) to help maintain the cells as pluripotent stem cells. This protocol has been optimized for the ES-D3 cell line. - [Read Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Mouse Embryonic Fibroblasts Protocols

Serum Testing for Mammalian Cell Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4348

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

This protocol describes a method for testing of new serum lots prior to use in cell culture. Serum lots vary considerably in their ability to support cell growth, and some lots even contain toxic or growth-inhibitory compounds. It is advantageous to test serum lots and purchase in large volume, both for cost benefit. - [Read Serum Testing for Mammalian Cell Culture Protocol]

Articles (1-2 of 2)

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

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