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KnockOut™ Serum Replacement (KSR)

Quick Facts

FeatureInformation
Supplement TypeSerum replacement
CompositionProprietary defined formulation
Typical Working Concentration15-20% (v/v)
Primary FunctionMaintains pluripotency while reducing spontaneous differentiation
Primary ApplicationESC and iPSC culture

Overview

KnockOut™ Serum Replacement (KSR) is a chemically defined alternative to fetal bovine serum (FBS) developed specifically for pluripotent stem cell culture. Compared with serum, KSR provides a more controlled culture environment, minimizes batch-to-batch variability, and reduces spontaneous differentiation.

KSR is most commonly used with KnockOut™ DMEM for the maintenance, expansion, reprogramming, and genome editing of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs).

Composition

Although the complete formulation is proprietary, KSR contains a defined mixture of:

ComponentFunction
AlbuminCarrier protein and osmotic support
LipidsMembrane synthesis
Amino acidsProtein synthesis
VitaminsCellular metabolism
AntioxidantsProtection against oxidative stress
Trace elementsEnzyme cofactors

Biological Role

KSR supports:

  • Pluripotent stem cell self-renewal
  • Colony formation
  • Cell survival
  • Controlled proliferation
  • Genome stability
  • Reduced spontaneous differentiation

Typical Applications

KSR is routinely used for:

  • Human embryonic stem cells (hESCs)
  • Mouse embryonic stem cells (mESCs)
  • Induced pluripotent stem cells (iPSCs)
  • Stem cell reprogramming
  • CRISPR genome editing
  • Clonal expansion
  • Developmental biology
  • Directed differentiation protocols

Recommended Use

ParameterRecommendation
Working Concentration15-20% (v/v)
Mouse ESCsTypically 15%
Human ESCs/iPSCsTypically 20%

Always optimize the concentration according to the published protocol or cell line recommendations.

Advantages

  • Defined alternative to FBS
  • Excellent batch-to-batch consistency
  • Supports long-term pluripotency
  • Reduces spontaneous differentiation
  • Highly reproducible
  • Well established in stem cell research

Limitations

  • Proprietary formulation
  • More expensive than FBS
  • Primarily intended for pluripotent stem cells
  • Increasingly replaced by fully defined feeder-free media (e.g., Essential 8, StemFlex) for some applications

Comparison with Related Supplements

SupplementPrimary Function
KSRDefined serum replacement for pluripotent stem cells
Fetal Bovine Serum (FBS)Undefined serum supplement for general cell culture
Essential 8 / StemFlexFully defined media for feeder-free pluripotent stem cell culture

References

  • Gibco™ KnockOut™ Serum Replacement Product Information.
  • Thomson JA et al. Embryonic stem cell lines derived from human blastocysts. Science (1998).
  • Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell (2006).
  • Freshney RI. Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications.
  • ATCC Animal Cell Culture Guide.