Background

The use of mouse ES (mES) cells to generate transgenic animals has revolutionized the study of gene action in vivo, and has provided animal models for studying the pathophysiology of human genetic disease and therapies thereof. Since the discovery that inherited mitochondrial DNA (mtDNA) mutations can cause age-related degenerative diseases (1), it has been realized that inherited mtDNA mutations are also associated with human adaptation to different environments, a broad spectrum of...

Abbreviations

Chemically defined medium complementary DNA (sequence) cES-cell medium cynomolgus ES (cells) colony forming unit CFU-erythroid CFU-granulocyte 2'3'-cyclic nucleotide 3'-phosphodiesterase DMSO, acetamide and propylene glycol (freezing medium) dibutyryl-cyclic AMP direct current diethyl pyrocarbonate Dil-Ac-LDL Dil-acetylated-low density lipoprotein DMEM Dulbecco's modified Eagle's medium DPX di-n-butyl phthalate xylene (mountant) ECs endothelial cells (Chapter 11) EDTA ethylenediaminetetraacetic...

Animal transplantation models

For the validation of in vitro generated, mES cell-derivatives as being pheno-typically functional and having regenerative capacity, animal disease and transplantation models are essential. These disease models, ideally, should demonstrate similar pathological mechanisms and properties to human disease syndromes and may be either induced experimentally or naturally occurring. Several significant murine i.e. mouse and rat models of pancreatic and liver diseases are now established, and have been...

Differentiation of mES cells into muscle cell lineages

When mES cells are induced to form aggregates in vitro, in the absence of those self-renewal signals normally provided by mitotically inactivated feeder layers or LIF, cell differentiation is initiated see Chapter 5 . If aggregates are allowed to form spontaneously in suspension by seeding a culture onto a non-adhesive substratum typically, a bacteriological-grade Petri dish , the number of cells incorporated into each aggregate is variable. But this number can be controlled through the use of...

Methods for the derivation of mES cell lines

The classical and standard method for deriving mES cell lines is to culture intact day 3.5 p.c blastocysts on feeder layers for 2-5 d during which time the blastocysts hatch and attach to the substratum , and then to collect, dissociate, and subculture the cells of the explanted ICM Protocol 7, Method A . With blastocysts from strain 129 or C57BL 6 mice, lines should be obtained efficiently by this technique without recourse to other methods. Otherwise, improved efficiencies may be achieved by...