Mouse Mesenchymal Stem cells: Advanced pre-screening tools for Toxicological Studies
                     Mouse Mesenchymal 
Stem cells: Advanced 
pre-screening tools for 
Toxicological Studies
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Introduction
• Overview of Mesenchymal Stem cells and their unique properties
Mesenchymal stem cells are multipotent stem cells and are highly acknowledged for their 
differentiation abilities into multiple cell types; which make them an ideal choice for organ-
specific drug toxicity at a cellular level. They have rapid multiplication abilities, enabling them to 
be exposed to multiple drug candidates in parallel for comparative analysis.
• How can they be a relevant choice for toxicological studies
The mesenchymal stem cells exhibit a rapid response rate upon exposure to toxins at both 
genotypic and phenotypic levels. This ability of MSCs is helpful for the early identification of 
drug-induced toxicities at cellular and molecular levels, through various assays.
• Importance of in vitro screening in toxicological studies
In vitro studies are an ethical approach in toxicological studies, bypassing the direct recruitment 
of animals in scientific research. Due to the easy expansion capacities of certain cells like MSCs, 
in vitro screening studies are cost-effective solutions, saving time and resources.
Study Rationale
• Why should one select MSC derived from C57 and BalbC stains only
In toxicological research, C57 and BalbC are two of the most widely used inbred mouse strains. 
They are highly preferred due to their genetic similarity with humans and reproducibility.
• Limitations associated with in vivo toxicological studies
Since these animals have relatively short lifespans, long-term toxicity studies may be a 
challenging task, whereas mesenchymal stem cells derived from these animals can be cryo-
stored for years to mimic the in vivo environment. Moreover, ethical concerns can complicate the 
in vivo animal toxicity studies due to stringent regulations can significantly be challenging due to 
cost in research setups.
The MSCs derived from these strains
• Advantages over traditional 2D or other cell line-related models
• Other advantages include a reduction in animal usage and alignment with the 3Rs principles 
(Replacement, Reduction, and Refinement).
Applications in Prescreening Toxicological 
PSrteudidctiiengs cytotoxicity, genotoxicity, and mitochondrial 
dysfunctions
1.  It is possible to assess drug-induced cytotoxicity on MSCs derived from the 
proposed animals through evaluation of direct impact on cell viability and measuring 
secretions of certain hormones like LDH.
2.  It is possible to assess toxicity at genotypic levels by assessing mutagenicity 
caused by test compounds and visualize genetic disruptions at telomeric levels 
through comet assays.
3.  With the help of MSCs it is also possible to evaluate the role of a test compound 
on mitochondrial dysfunction through investigation of mitochondrial activity.
Testing dose-dependent responses for candidate compounds
1.  The in vitro-grown MSCs can be the ideal choice for 
performing dose-dependent studies using a systemic 
titration of drug compounds, due to their high 
expansion potential and low doubling time.
2.  Due to the higher sensitivity of MSCs towards the 
assay conditions, the cellular behavior can be easily 
determined for survival, proliferation, and functionality 
at varied concentrations of a test compound.
3.  With the great differentiation capacity of MSCs to 
different organs, the safety margins and therapeutic 
efficacy of a test compound can be checked at the 
cellular level.
Modeling immune modulation effects and cytokine 
profiling
1.  MSCs are well-known for their immune-
modulatory functions, thus they can be 
the ideal choice to examine how a test 
compound can affect cellular behavior 
through examination of an inflammatory 
response.
2.  The immunomodulatory ability of a 
test compound can be assessed through 
the evaluation of secretary inflammatory 
and anti-inflammatory growth factors.
Experimental workflow
• Cell isolation and characterization from the specified strains
1.  Get assay-ready MSCs from C57 and/or BalbC mouse strains of your choice; which 
are well characterized for their surface markers, viability, and growth.
2.  Further ensure that the cells are stabilized for proliferation rates and normal 
karyotyping to ensure consistency of results for downstream processing.
•    Steps in pre-screening studies like
1.  cell viability assays including MTT, LDH release, etc.
2.  Evaluation of cellular stress and oxidative damage
3.  Multi-lineage differentiation capacity post-toxin exposure
• Data interpretation and decision making
1.  Result in compilation and further analysis
Future directions
• Integration with other high-throughput screening platforms
⚬ The MSCs can serve as a good choice for employing organ-on-chip models to 
trigger responses at organ levels, and also capture inter-organ 
communication effects.
• Incorporating OMICS for deeper insights
⚬ These cells can be easily expanded at a larger scale for other downstream 
applications like metabolomics and proteomics to understand metabolic and 
genetic shifts upon exposure to a test compound.
• Extending applications to humanized toxicology models
⚬ As MSCs are also present in every species, it is also possible to adapt the 
platform for human-derived MSCs to create physiologically relevant models 
for deeper understanding.
Conclusion
Contribution to safer and more efficient drug development pipelines
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