Related Experiment Video
Updated: Aug 9, 2026
A Strategy to Identify Compounds that Affect Cell Growth and Survival in Cultured Mammalian Cells at Low-to-Moderate Throughput
Published on: September 22, 2019
Cell detachment and cloning efficiency as parameters for cytotoxicity
Abstract:
Cell detachment and cloning efficiency of Baby Hamster Kidney cells (BHK-21 C13) were used as parameters to quantify cytotoxicity in vitro of 3 endogenous chemicals (glutathione, L-methionine, L-cysteine HCl) and 4 organotin compounds (tributyltinoxide, tributyltinchloride, tetrabutyltin, tetraphenyltin). IC50 values (inhibitory concentration at which the cloning efficiency was reduced to 50%) were estimated to be larger than 10(-3) M for all 3 endogenous substances, which served as a calibration of the cell culture system for non-toxic chemicals. For the 2 tributyltin salts the IC50 values were estimated to be near 10(-6) M and for the 2 tetraalkyltin compounds near 10(-5) M. The estimated CD50 values (concentration at which 50% of the cells detach) were at least twice as large as the corresponding IC50 values for all 7 chemicals tested. For the toxic tributyltin salts the cell detachment assay was 30-60 times less sensitive than the cloning efficiency assay. However, both assays rank all compounds tested in the same sequence of toxicity as that known from in vivo studies.
Insights
This study evaluated the cytotoxicity of organotin compounds using Baby Hamster Kidney cells. Organotins showed higher toxicity than endogenous substances, with cell detachment assays correlating with cloning efficiency assays for toxicity ranking.
Area of Science:
- Toxicology
- Cell Biology
- Environmental Chemistry
Background:
- Cytotoxicity assessment is crucial for evaluating chemical safety.
- Organotin compounds are known environmental pollutants with varying toxicity profiles.
- Standardized in vitro assays are needed for reliable toxicity quantification.
Purpose of the Study:
- To quantify the in vitro cytotoxicity of specific endogenous chemicals and organotin compounds.
- To compare the sensitivity of cell detachment and cloning efficiency assays for cytotoxicity.
- To correlate in vitro toxicity findings with known in vivo toxicity data.
Main Methods:
- Utilized Baby Hamster Kidney (BHK-21 C13) cells for cytotoxicity testing.
- Assessed cytotoxicity by measuring cell detachment and cloning efficiency.
- Determined IC50 (inhibitory concentration for 50% cloning efficiency) and CD50 (concentration for 50% cell detachment) values.
Main Results:
- Endogenous substances (glutathione, L-methionine, L-cysteine HCl) exhibited low cytotoxicity (IC50 > 10(-3) M).
- Organotin compounds showed significant cytotoxicity: tributyltin salts (IC50 ~ 10(-6) M) and tetraalkyltins (IC50 ~ 10(-5) M).
- CD50 values were consistently higher than IC50 values; cell detachment assay was less sensitive for toxic tributyltins.
Conclusions:
- Both cell detachment and cloning efficiency assays effectively rank organotin compound toxicity.
- In vitro results align with known in vivo toxicity sequences for the tested chemicals.
- The study validates these assays for assessing organotin compound cytotoxicity and environmental risk.
