Cell culture is a powerful and versatile technique that enables researchers to explore the intricacies of cell biology, understand disease processes, develop novel therapeutics, and advance various fields of biomedical research.

At Visikol we specialize in many different advanced cell culture methods, one of them being 3D cell culture models. 3D cell culture is an innovative and advanced approach in in vitro research that aims to mimic the complexity and physiological relevance of the human body more accurately than traditional 2D cell culture. The three-dimensional nature of 3D cell culture provides a better understanding of biological processes; it allows for the study of cell migration, invasion, angiogenesis, and cell signaling in a more realistic context. This helps elucidate the underlying mechanisms of disease progression and cellular interactions.

Cells grown in 3D environments can form complex structures, including cell clusters, spheroids, organoids, and tissue-like structures, which closely resemble the organization and function of tissues in vivo. Spheroids mimic the cellular heterogeneity and cellular interactions found within solid tumors, and as a result provide a realistic tumor-like physiology that allows for the study of drug resistance and facilitates the development of targeted therapies. Researchers can expose spheroids to various compounds and evaluate their effects on tumor growth, proliferation, viability, and other relevant endpoints. The 3D nature of spheroids allows for a more accurate prediction of drug responses and can help identify potential drug candidates with improved efficacy against tumors.

In 3D cell culture, cells exhibit improved cellular function and phenotype as compared to 2D cell culture. Cells grown in 3D environments often display enhanced differentiation, polarization, and gene expression patterns that more closely resemble their in vivo counterparts. Because these 3D structures can mimic specific organs or tissues, researchers can create more accurate models of diseases such as cancer, neurodegenerative disorders, and organ-specific diseases. These models allow for the study of disease progression, drug responses, and the screening of potential therapeutic compounds.

Assessing the potential toxicity of drugs is crucial in drug development. The 3D environment better replicates the in vivo conditions, including drug diffusion, metabolism, and cellular responses. This enables more accurate assessment of drug-induced toxicity, drug efficacy, and pharmacokinetics, reducing the reliance on animal models. The use of 3D cell culture models bridges the gap between traditional in vitro experiments and in vivo studies. The more physiologically relevant environment provided by 3D culture enhances the translatability of research findings, making them more applicable to human biology. This accelerates the translation of basic research discoveries into clinical applications and therapeutic interventions.

Incorporating 3D cell culture into pharmacology research can lead to more accurate and efficient drug development processes, ultimately improving drug safety and efficacy. Contact us today to work with our expert team of scientists!

When working on drug discovery and assay development, scientists often use models that can help mimic the effects of the drugs of interest. This can range from 2D models in a plate, trans-well models with cell suspension and 3D models such as spheroids or organoids. Spheroids are a miniature unit that mimics what a tumor is and is quick and easy to grow compared to an organoid, which is a 3D structure that has grown to replicate the size and the shape of the organ of interest.

It is useful to have spheroids and use them in various experiments. Cells take about 48 hours to form spheroids once plated into a ULA round bottom plate, however this is dependent on the cell line. As some cell structures are not all uniformly congruent, this can impose challenges in even distribution of drug dosing, varying penetration rates and structural deformities. This poses the need for an effective way to make spheroids to induce the effects the drug has on the cells.

Initial Plating

To ensure the correct usage of the cell in a spheroid format, it is encouraged to do an initial plating onto a ULA plate to encourage natural spheroid formation. An example of such case is that when specific cell lines are known to have non-symmetrical shapes. Some examples are of A549 cancer cell line, which has an elongated shape and is not known to form round spheroids. However, doing an initial run on the cells for 48 hours to determine the final shape of the spheroids is deemed helpful when planning to run tests on them.

Using spheroids in drug discovery research is like managing a small-scale cancer in an organism. Although utilizing spheroids is in vitro, this tool is effective in giving a better idea on how to move forward with dosing, assay procedures and implementing data on patients later down the road.

Visikol incorporates spheroids as an essential part of drug discovery, and it allows clients to understand the physiology of the cells of interest as they come together to mimic a miniscule version of a tumor. Using various techniques and previous experience handling cells to form spheroids, Visikol can provide an in-depth service in cell culture assays as well as offering many other types of services. To learn more about Visikol’s assays and various testing services, please reach out.

With the increased adoption of holistic health approaches and a stronger consciousness of how the environment impacts our health, an important application of in vitro studies arises. A growing population has become concerned with how extraneous chemicals interrupt endocrine function, and recent advances in cell culture techniques allow researchers to effectively investigate this question. Visikol is a leader in the cell culture industry, utilizing patented clearing technologies and robust 3D models to evaluate the functioning of major organ systems in both healthy and diseased states. One of the body’s most vital systems – the hypothalamic-pituitary-thyroid (HPT) axis – is of particular research interest when it comes to endocrine perturbations because it is fundamental to metabolic equilibrium, growth, and cognitive development.

Plant protection products (PPPs) are particularly prevalent environmental stressors, but the extent to which they interrupt thyroid hormones (TH) is a question that remains for regulatory authorities.

The Method

In a recent publication entitled, “A Microfluidic Thyroid-Liver Platform to Assess Chemical Safety in Humans,” researchers investigated human-relevant interferences of thyroid gland function through the HPT axis – or more specifically, TH catabolism through hepatic enzyme induction. To address the limitations of standard human in-vitro assays, researchers utilized human 3D thyroid and liver organoids on a micro physiological organ chip to evaluate the effects of methimazole (a reference thyroid peroxidase (TPO) inhibitor) over the course of 21 days. With the help of Visikol’s HISTO-M®, samples were cleared to allow for characterization of 3D thyroid model morphology and morphological changes, cAMP accumulation, thyroid hormone secretion, and TPO inhibition. Additional characterization was performed on the liver spheroids to understand TH catabolism, glucuronidated and sulfated thyroxine (gT4/sT4) metabolite formation, and activation of nuclear receptors. Lastly, the functionality of these two 3D models together was assessed on a perfused multi-organ chip platform (the Chip2) to simulate the in vivo interaction between these two organ systems.

Overall, 3D cell culture models allow us to assess the human relevance of safety-related findings in animal models with respect to thyroid toxicity. This research is essential to the development of new PPPs and represents scientifically informed advancement toward a safer, healthier environment. Visikol’s attention to the development of highly useful 3D models is an essential element not only to this research but also to our understanding of how complex organ systems work. Please reach out to explore what Visikol can do for you.

Thyroid hormones are important for regulating metabolism, development, and a variety of other body functions. While human in vitro assays are used to elucidate human-relevant interferences with thyroid gland function, they do not adequately reflect the complex TH biology and related liver-thyroid relevance; in order to address this issue, Kuhnlenz et al. developed human 3D thyroid and liver organoids, which stimulate TH biosynthesis and hepatic TH catabolism. In this study, the liver spheroids model made from HepaRG/hepatic stellate cells (HSteC) were capable of simulating TH catabolism by producing the compounds glucuronide conjugated H thyroxine (Gt4) and sulfate conjugated thyroxine 3 (St3). The ultimate goal of the study was to functionally co-cultivate pre-characterized thyroid and liver organ models on a commercially accessible multi-organ chip (MOC) technology due to the complexity of TH homeostasis, which is often impacted by both direct and indirect thyroid gland deficiencies. The co-culture setup was undertaken in order to support the functions of single-tissue organ-chip tissues for over several days.

Samples were cleared in Visikol Histo-M for histological analysis, particularly for whole tissue 3D staining, which aided in the characterization of the 3D thyroid model. Thyroid model cultures exposed to different TSH concentrations showed significantly higher cAMP levels after exposure to 1 and 10 mIU/ml TSH, indicating that the 3D thyroid model is responsive to an external TSH-stimulus, authors state. Moreover, the models from four human donors were pre-cultured for three days before being treated for four days with non-cytotoxic concentrations of Methimazole (MMI), a reference Thyroid peroxidase (TPO) inhibitor. After 2 and 4 days of treatment, triiodothyronine (T3) secretion levels in culture supernatants were measured and normalized to DMSO-solvent controls; it was found that the minimal inhibitory concentration of 1 M MMI reliably inhibited T3-secretion, but after several days, it was found that 10 M MMI had the greatest inhibitory effect, suppressing T3-secretion continuously. These results suggest that the established 3D thyroid model can detect immediate TH perturbations on T3 synthesis levels. Further analysis revealed that the established liver spheroids model maintains meaningful features which contribute to an in-vivo like TH homeostasis.

Overall, the established human 3D thyroid model is the first in vitro thyroid model that can be integrated into a perfused organ-chip system with viability and TSH-dependent T3 secretion over 21 days; however, further model improvement must be considered if one wishes to follow physiological T4/T3 ratio, for instance, by understanding and observing DIO1/2 activity in the system; DIO1, the only gene expression currently under investigation, did not significantly differ from levels of expression found in primary tissues, Kuhnlenz et al. state.

Visikol offers a wide range of tissue clearing reagents for transparentizing biological tissues and cell models, as well as advanced drug discovery solutions such as 3D cell culture assays  and  tissue imaging high content screening, AI solutions for histological analysis of tissue sections, and much more. To learn more about Visikol’s services, please contact us.

The realm of 3D cell models has grown exponentially in a short period of time, gaining traction with researchers due to their dynamic and complex nature. Visikol is a leader in the cell culture industry, utilizing robust 3D models to evaluate the functioning of major organ systems in both healthy and diseased states. One highly useful model, the spheroid, has recently been used to study the largest organ in the human body – the skin. Specific proteins, such as carnosine, have been shown to prevent the visible signs of aging and damage related to UV exposure, and 3D models prove to be a useful tool for in-depth tissue characterization.

Skin cancer related to UV damage is a commonly encountered concern today but understanding our skin in its healthy state is a fundamental step toward identifying effective treatments.

In a recent publication entitled, “Oxidative Stress Modulation by Carnosine in Scaffold Free Human Dermis Spheroids Model: A Proteomic Study,” researchers investigated the mechanism of action of carnosine – an endogenous β-alanyl-L-histidine dipeptide – on human skin proteome. Carnosine is a known antioxidant and carbonyl scavenger, making it a strong defender against aging of the skin. In this study, a spheroid model was adopted from human dermal fibroblasts and combined with high-resolution mass spectrometry to characterize the protein expression during long-term culture upon carnosine treatment. At 14 days of culture, with 7 days of culture time as a benchmark, carnosine was found to activate the functionality of mitochondria – specifically the upregulation of functional modules related to oxidative stress. To evaluate localization and modification during culture time, immunofluorescence for collagen type III was analyzed with the help of Visikol’s Histo-M Starter Kit®. Upon tissue clearing and signal quantification, proteins were extracted and analyzed via high-resolution LC-MS/MS, then statistically evaluated.

Overall, 3D models are highly phenotypically relevant to human tissues in a way that can advance science toward a more complete approach to combating disease. The diverse range of products and services Visikol has to offer can pave the way for personalized care therapies and beyond. Please reach out to explore what Visikol can do for you.