CELL LINES
Contributor Information
- Name Annemieke Aartsma-Rus
- Institute Leiden University and Leiden University Medical Center
Tool Details
- Tool name: hDMD/mdx ES cell line
- Alternate names: Dystrophin, Muscular Dystrophy, Duchenne And Becker Types, DXS164, DXS26, DXS23, DXS239, DXS268, DXS269, DXS27, DXS272
- Tool type: Cell Lines
- Parental cell line: Blastocysts were cultured to generate ES cell lines
- Organism: Human
- Tissue: Embryo
- Disease: Duchenne Muscular dystrophy
- Model: Stem Cells
- Conditional: No
- Description: Duchenne muscular dystrophy (DMD) is a muscle-wasting disease in which muscle is continuously damaged, resulting in loss of muscle tissue and function. Antisense-mediated exon skipping is a promising therapeutic approach for DMD which uses sequence specific antisense oligonucleotides (AONs) to reframe disrupted dystrophin transcripts. As AONs function in a sequence specific manner, human specific AONs cannot be tested in the current mdx mouse model for DMD as it carries a mutation in the murine Dmd gene. In order to model the human disease more accurately we generated an ES mouse cell line carrying the complete human DMD gene integrated in the mouse genome on an mdx background. This cell line was used to generate the hDMD/mdx mouse (Cat No:154187)
- Research area: Cell biology; Drug development
- Production details: Blastocysts were isolated from time mated hDMD male mice and super ovulated mdx female mice. These blastocysts were layered on murine embryonic fibroblast (MEF) feeder cells in a well of a 24-well plate, in knockout DMEM supplemented with 2 mM L-glutamine, 1 mM sodium pyruvate, non-essential amino acids, 50 units/ml of penicillin as well as streptomycin, 1000 units/ml of LIF) and 15% knockout serum replacement. Usually after 6 days blastocysts had hatched and a small colony of cells had formed. These were trypsin digested and cells were placed in a new MEF coated well of a 24-well plate. To stop trypsin activity cells were cultured overnight in ES medium supplemented with 10% fetal bovine serum. The next morning medium was replaced by ES medium with knock serum replacement. These steps were repeated several times till sufficient ES cells were available for freezing down and analysis cultured to generate ES cell lines
- For Research Use Only
Target Details
- Target: DMD
Application Details
Handling
- Format: Frozen
- Growth medium: ES medium with 15% knock serum replacement
- Shipping conditions: Dry ice
Documentation
Related Tools
References
- • Veltrop et al. 2013. PLoS Curr. 5:. PMID: 24057032
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