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mouse embryonic stem cell research
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Mini-Review
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for undifferentiated mESCs
for in vitro differentiation
mESC-screened support products

background
Mouse embryonic stem cells (mESCs) are pluripotent cells derived from the inner cell mass of day 3.5 blastocysts. They can be maintained in vitro for extended periods without loss of their capacity to contribute to all cell lineages when reimplanted back into a blastocyst.1,2 The pluripotency of mESCs, combined with their ease of genetic manipulation and selection, has revolutionized gene function studies in vivo via the generation of transgenic, chimeric and knockout mice.3-8 Mouse ESCs are maintained in an undifferentiated state in vitro by precise and defined culture conditions in the presence of leukemia inhibitory factor (LIF). Upon withdrawal of LIF, mESCs differentiate into complex structures called embryoid bodies (EBs) that contain cells from each of the three germ layers: endoderm, ectoderm, and mesoderm. These cells can subsequently be induced to differentiate into a variety of cell types9,10 including pancreatic,11neural,12-17 hematopoietic,18-20 cardiomyocytes,21-24 adipocytes,25 epithelial,26 endothelial,27-31 lymphoid cells,32-36 and osteoblasts.37 Mouse ESC in vitro differentiation provides a unique and powerful system to examine cellular and molecular processes, perform drug screening, and to investigate potential tissue engineering and cellular therapy applications.

For more information, refer to the Mini-Review on mESCs.


reagents available for:

  • Growth and maintenance of germline competent mESCs for the generation of chimeric and transgenic mice
  • Growth and maintenance of undifferentiated mESs for in vitro differentiation to hematopoietic, pancreatic, endothelial and neural lineages
  • Formation of EBs in primary methylcellulose cultures
  • Formation of EBs in liquid suspension cultures
  • Disaggregation of EBs
  • Cytokine-directed generation of hematopoietic colonies in methylcellulose cultures
  • Production of insulin-secreting pancreatic islet-like clusters
  • Production of a variety of neural cell types including neurons, astrocytes and oligodendrocytes

quality control

All ES-Cult® reagents are screened and extensively pre-tested for optimal mESC performance.

All lots of serum are performance tested with an mESC line to ensure:

  • Optimum growth rates
  • No toxicity even at high (30%) serum concentrations
  • High cloning efficiency at low cell densities
  • Maintenance of undifferentiated mESC colony morphology (FBS; Catalog #06902/06952)
  • The ability to maintain undifferentiated mESCs for generation of chimeric and transgenic mice (FBS; Catalog #06902/06952)
  • Maintenance of a high hematopoietic differentiation potential (FBS; Catalog #06902/06952)
  • Efficient two-step in vitro differentiation into hematopoietic colonies (FBS; Catalog #06900/06950)
  • Efficient embryoid body formation in liquid suspension cultures (FBS; Catalog #06905/06955)

ES-Cult® methylcellulose-based media is extensively pre-tested to ensure it supports efficient generation of embryoid bodies and hematopoietic progenitors in a two-step in vitro differentiation assay.

All other ES-Cult® media, supplements and accessories are carefully pre-screened in mESC maintenance and/or differentiation assays.


applications

  • Analysis of gene expression patterns during embryonic development and differentiation
  • Provides ideal model systems for the generation of specifc cell types for potential therapeutic approaches
  • Creation of genetically modified mESCs (knockout, transgenic, antisense) for functional studies
  • Evaluation of the effects of external stimuli on developmental processes
  • Identification of novel developmentally regulated genes using gene trap, degenerate PCR, or RNA subtraction techniques


 

FOR RESEARCH USE ONLY

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