Quick Facts
| Feature | Information |
|---|
| Medium Type | Specialized basal medium |
| Parent Medium | Modified DMEM |
| Best For | Pluripotent stem cell culture |
| Common Applications | ESCs, iPSCs, reprogramming, genome editing, stem cell banking |
| Common Supplements | KnockOut™ Serum Replacement (KSR), NEAA, GlutaMAX™, β-Mercaptoethanol, FGF2 or LIF |
| Storage | 2-8°C, protected from light |
Overview
KnockOut™ DMEM is a modified formulation of DMEM developed specifically for embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). When combined with KnockOut™ Serum Replacement (KSR), it provides a defined culture environment that supports long-term self-renewal while reducing spontaneous differentiation.
Although many laboratories now use feeder-free commercial media, KnockOut™ DMEM remains a standard basal medium for feeder-based pluripotent stem cell culture, cellular reprogramming, and genome editing workflows.
Key Features
- Optimized for pluripotent stem cell maintenance
- Supports long-term self-renewal
- Reduces spontaneous differentiation
- Compatible with feeder-based and serum-free culture systems
- Widely used in stem cell reprogramming
- Suitable for CRISPR clone recovery and stem cell banking
Typical Supplements
| Supplement | Purpose |
|---|
| KnockOut™ Serum Replacement (KSR) | Serum replacement |
| GlutaMAX™ or L-Glutamine | Nitrogen metabolism |
| Non-Essential Amino Acids (NEAA) | Cell growth |
| β-Mercaptoethanol | Reduces oxidative stress |
| FGF2 | Human PSC maintenance |
| Leukemia Inhibitory Factor (LIF) | Mouse ESC maintenance |
| Penicillin-Streptomycin | Antibiotic |
Common Applications
Pluripotent Stem Cell Culture
- Human embryonic stem cells (hESCs)
- Mouse embryonic stem cells (mESCs)
- Human iPSCs
- Mouse iPSCs
Stem Cell Engineering
- Cellular reprogramming
- CRISPR genome editing
- Clone recovery
- Stem cell banking
Research Applications
- Developmental biology
- Regenerative medicine
- Pluripotency studies
- Disease modeling
Advantages
- Optimized for pluripotent stem cells
- Supports stable self-renewal
- Reduces spontaneous differentiation
- Compatible with feeder-based culture systems
- Well established in stem cell research
- Suitable for genome engineering workflows
Limitations
- Not intended for routine differentiated cell culture
- Requires multiple supplements
- More expensive than conventional DMEM
- Increasingly replaced by fully defined feeder-free media in some applications
Comparison with Related Media
| Medium | Best Suited For |
|---|
| KnockOut™ DMEM | ESCs, iPSCs, feeder-based pluripotent stem cell culture |
| DMEM | General mammalian cell culture |
| DMEM/F-12 | Stem cells, primary cells, organoids |
| Essential 8 (E8) | Chemically defined feeder-free pluripotent stem cell culture |
Best Practices
- Prepare fresh complete medium with appropriate supplements.
- Feed pluripotent stem cells daily unless following an alternative validated protocol.
- Regularly monitor colonies for spontaneous differentiation.
- Maintain low passage stocks whenever possible.
- Periodically confirm pluripotency using established molecular markers.
References
- Thermo Fisher Scientific. Gibco™ KnockOut™ DMEM Product Information.
- Thomson JA et al. Science (1998).
- Takahashi K, Yamanaka S. Cell (2006).
- ATCC. Animal Cell Culture Guide.
- Freshney RI. Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications.