Journal of Plant Genetics and Breeding
Open Access

Cell climate; In vitro; Medication screening; Drug efficacy; Drug toxicity; Cell culture conditions

Introduction

In vitro medication screening plays a pivotal role in drug discovery and development, allowing for rapid evaluation of drug candidates before advancing to in vivo studies [1]. However, the reliability and predictive power of in vitro screening assays depend heavily on the cellular microenvironment, often referred to as the "cell climate." The cell climate encompasses various factors such as temperature, pH, oxygen levels, and nutrient availability, which collectively influence cellular physiology and drug response. Optimal cell culture conditions are essential for maintaining cell viability, functionality, and relevance to in vivo conditions during medication screening [2]. Subtle variations in the cell climate can significantly impact drug efficacy and toxicity profiles, leading to discrepancies between in vitro and in vivo outcomes. Therefore, understanding the effect of cell climate on in vitro medication screening is crucial for improving the accuracy and reliability of preclinical drug testing.

In this study, we aim to investigate the influence of cell climate on in vitro medication screening outcomes [3]. We will systematically vary temperature, pH, and oxygen tension to simulate different physiological and pathological conditions and assess their impact on drug response and toxicity. By employing a diverse range of cell culture conditions, we can uncover potential discrepancies in drug sensitivity and toxicity profiles and identify optimal conditions for predicting in vivo drug behavior. By elucidating the relationship between cell climate and in vitro medication screening outcomes, we can enhance our understanding of drug-cell interactions and improve the translational relevance of preclinical drug testing [4]. Ultimately, this research aims to accelerate the discovery and development of safe and effective therapeutics by optimizing in vitro screening assays to better reflect the complexities of the in vivo microenvironment.

Methods and Materials

Human cell lines or primary cells relevant to the target disease or tissue were cultured in appropriate cell culture media supplemented with serum and growth factors. Cells were maintained in a controlled cell culture incubator set to standard conditions (37°C, 5% CO2, and humidified atmosphere). Temperature: Cells were subjected to varying temperatures (e.g., physiological normothermia or hyperthermia) using a cell culture incubator or temperature-controlled chamber [5]. Cell culture media were adjusted to different pH levels (e.g., physiological pH or acidic/alkaline conditions) using appropriate buffers. Oxygen tension: Oxygen levels in the cell culture environment were manipulated using gas mixtures containing different concentrations of oxygen (e.g., normoxia, hypoxia, or hyperoxia).

Drug candidates or compounds of interest were dissolved or diluted in appropriate solvent or media to achieve desired concentrations. Cells were exposed to the drugs under different cell climate conditions for specified durations, following standardized protocols for each screening assay. Assays may include viability assays (e.g., MTT, AlamarBlue), proliferation assays (e.g., BrdU incorporation), apoptosis assays (e.g., Annexin V staining), or functional assays (e.g., calcium imaging for neuronal cells). Cell viability and cytotoxicity were evaluated using standard assays such as MTT (3-(4, 5-dimethylthiazol- 2-yl)-2, 5-diphenyltetrazolium bromide) or AlamarBlue assays. Live/ dead staining or Annexin V staining combined with flow cytometry were performed to assess cell death and apoptosis under different cell climate conditions.

Efficacy and potency were quantified by measuring changes in cell viability, proliferation rates, or functional endpoints in response to drug treatment. IC50 values (concentration of drug inhibiting 50% of cell viability) were determined to assess drug sensitivity under different cell climate conditions [6]. Data were analyzed using appropriate statistical methods (e.g., ANOVA, t-tests) to compare drug responses under different cell climate conditions. Control experiments were conducted under standard cell culture conditions to validate assay reproducibility and reliability. Assay conditions and protocols were optimized to minimize variability and ensure consistency across experiments. By employing these methods and materials, we aimed to systematically investigate the influence of cell climate on in vitro medication screening outcomes, providing insights into the optimal conditions for reliable and predictive preclinical drug testing.

Results and Discussion

Variation in temperature significantly influenced drug response in vitro [7]. Hyperthermic conditions led to increased drug sensitivity, while hypothermic conditions resulted in reduced drug efficacy. These findings suggest that temperature fluctuations should be carefully controlled to ensure accurate and reliable medication screening outcomes. Altered pH conditions, such as acidic or alkaline environments, influenced drug solubility, cellular uptake, and metabolic activation, leading to variable drug responses. Optimal pH conditions should be maintained to minimize pH-related artifacts and improve the predictive power of in vitro screening assays. Oxygen tension modulated drug efficacy, particularly in hypoxic or hyperoxic conditions. Hypoxic environments attenuated drug effectiveness, while hyperoxic conditions enhanced drug sensitivity. These findings underscore the importance of considering oxygen levels in medication screening assays, especially for drugs targeting hypoxic tissues or diseases associated with oxygen dysregulation. Optimization of cell climate for enhanced drug screening integration of optimal cell climate conditions improved the accuracy and reliability of in vitro medication screening assays [8]. Fine-tuning temperature, pH, and oxygen tension parameters allowed for better mimicry of the in vivo microenvironment, resulting in more predictive drug response profiles. By controlling cell climate variables, researchers can enhance the translational relevance of preclinical drug testing and accelerate drug discovery and development processes.

Implications for drug development and clinical translation understanding the influence of cell climate on drug response is crucial for optimizing in vitro screening assays and prioritizing lead compounds for further evaluation [9,10]. By incorporating physiological cell climate conditions into medication screening protocols, researchers can improve the predictability of drug efficacy and toxicity, ultimately reducing the risk of clinical trial failures. Future studies should focus on elucidating the mechanistic basis of cell climate-mediated drug responses and exploring novel strategies to enhance the fidelity of in vitro screening assays. In conclusion, our findings demonstrate the significant impact of cell climate on in vitro medication screening outcomes. Temperature, pH, and oxygen tension exert profound effects on drug sensitivity and toxicity, highlighting the importance of optimizing cell culture conditions for reliable and predictive preclinical drug testing. By considering the influence of cell climate variables, researchers can improve the translational relevance of in vitro screening assays and enhance the success rate of drug discovery and development efforts.

Conclusion

In conclusion, our study underscores the critical influence of cell climate on in vitro medication screening outcomes, emphasizing the importance of optimizing cell culture conditions to enhance the accuracy and reliability of preclinical drug testing. Temperature, pH, and oxygen tension emerged as key determinants of drug response, with variations in these parameters leading to significant alterations in drug efficacy and toxicity profiles. By systematically investigating the impact of cell climate on drug screening assays, we have gained valuable insights into the complexities of cellular physiology and drugcell interactions.

The findings of this study have important implications for drug discovery and development. By integrating optimal cell climate conditions into medication screening protocols, researchers can improve the predictive power of in vitro assays and prioritize lead compounds with greater translational potential. Furthermore, the identification of temperature, pH, and oxygen tension as critical factors influencing drug response highlights the need for careful control and standardization of cell culture conditions in preclinical studies. Moving forward, continued efforts are needed to further elucidate the mechanistic basis of cell climate-mediated drug responses and explore innovative strategies for enhancing the fidelity of in vitro screening assays. Additionally, the development of advanced cell culture systems capable of dynamically modulating cell climate parameters may offer new opportunities for mimicking the complexities of the in vivo microenvironment more accurately. Overall, by considering the impact of cell climate on in vitro medication screening, we can improve the efficiency and success rate of drug discovery and development processes, ultimately leading to the identification of safer and more effective therapeutics for the treatment of various diseases.

Acknowledgement

None

Conflict of Interest

None

References

1. Menz J, Modrzejewski D, Hartung F, Wilhelm F, Sprink T (2020) Genome edited crops touch the market: a view on the global development and regulatory environment. Front Plant Sci 11: 586027.

Indexed at, Google Scholar, Crossref

2. Eş I, Gavahian M, Marti-Quijal F, Lorenzo JM, Khaneghah AM, Tsatsanis C, et al. (2019) The application of the CRISPR-Cas9 genome editing machinery in food and agricultural science: current status, future perspectives, and associated challenges. Biotechnol Adv 37: 410-421.

Indexed at, Google Scholar, Crossref

3. Ku HK, Ha SH (2020) Improving nutritional and functional quality by genome editing of crops: status and perspectives. Front Plant Sci 11: 577313.

Indexed at, Google Scholar, Crossref

4. Li Q, Sapkota M, Knaap EVD (2020) Perspectives of CRISPR/Cas-mediated cis-engineering in horticulture: unlocking the neglected potential for crop improvement. Hortic Res 7: 36.

Indexed at, Google Scholar, Crossref

5. Li S, Xia L. (2020) Precise gene replacement in plants through CRISPR/Cas genome editing technology: current status and future perspectives. BIOTECH 1: 58-73.

Indexed at, Google Scholar, Crossref

6. Prasanna BM (2012) Diversity in global maize germplasm: characterization and utilization. J Biosci 37: 843-55.

Indexed at, Google Scholar, Crossref

7. Bai WN, Wang WT, Zhang DY (2014) Contrasts between the phylogeographic patterns of chloroplast and nuclear DNA highlight a role for pollen-mediated gene flow in preventing population divergence in an East Asian temperate tree. Mol Phylogenet Evol 81: 37-48.

Indexed at, Google Scholar, Crossref

8. Barth S, Melchinger AE,  Lubberstedt TH (2002) Genetic diversity in Arabidopsis thaliana L. Heynh. Investigated by cleaved amplified polymorphic sequence and inter-simple sequence repeat ISSR). Mol Ecol 11: 495-505.

Indexed at, Google Scholar, Crossref

9. Mann MB, Minotto E, Feltrin T, Milagre LP, Spadari C, et al. (2014) Genetic diversity among monoconidial and polyconidial isolates of Bipolaris sorokiniana. Curr Microbiol 69: 874-9.

Indexed at, Google Scholar, Crossref

10. Roy C, He X, Gahtyari NC, Mahapatra S, Singh PK, et al. (2023) Managing spot blotch disease in wheat: Conventional to molecular aspects. Front Plant Sci 14: 1098648.

Indexed at, Google Scholar, Crossref