Translational Insights for Reproducible Advanced Cell Systems
Cells are the building blocks of all organisms. Understanding the biology of cells and their interactions at the molecular level is the foundation of biomedical and life science research. Utilizing cells as models is an established approach for drug discovery, whereas their use as therapeutics is a rapidly growing approach.
More than 90% of therapeutic candidates fail to make it past phase III clinical trials, mainly due to their lack of efficacy. The use of 3D and stem cell-based models better recapitulate the pathobiology of diseases and offer in-depth predictive insights that can translate into positive clinical outcomes.
In recognition of this, the U.S. Food and Drug Administration (FDA) Modernization Act 2.0 allows scientists to now use cell-based models as alternatives to animal testing for drug applications.
Advanced cell models - such as organoids, spheroids and 2D stem cell-based models - are revolutionizing biomedical research and drug discovery. However, they come with several challenges:
Sartorius addresses these challenges and more with cutting-edge analytical solutions that streamline advanced cell model R&D workflows and provide translational insights that translate into clinical outcomes for patients in need.
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Traditional, 2D cell cultures utilizing human induced pluripotent (iPSCs) and mesenchymal stem cells (MSCs) have emerged as versatile and effective platforms not only for therapeutic purposes but also for disease modeling and drug discovery. iPSCs allow to produce quasi limitless supply of patient-specific somatic cells and the possibility to differentiate them into disease -relevant cell types enabling the study of human disease pathophysiology and the identification of new drug targets as well as testing of compounds in vitro.
Monitoring cell characteristics and analyzing of secreted proteins as well as assessment of gene expression is crucial to ensure the quality and reproducibility of the experiment, such us disease modeling, drug discovery and screening. Some parameters are tested throughout the culturing process including cell health and counts, and testing for microbial contaminations is recommended at several steps to ensure reliability and reproducibility of results. Sophisticated, quantitative analytical solutions which provide multi-parametric assays such Incucyte® Live-Cell Analysis and iQue®3 High-Throughput Cytometry platforms can greatly support the assessment of critical quality attributes.
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Creation of cell-based assays for downstream analysis and specific applications for the research area of interest such us disease modeling, drug discovery and compound screening.
Selected, clonal cells are further expanded utilizing growth factors and cytokines to obtain sufficient volumes for research and development models and downstream analytical purposes.Use the Incucyte® Live-Cell Analysis System to optimize and define cell culture regimes.
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Selection of the best cells or colonies based on specific markers. i.e. pluripotency to ensure the best cells are taken forward. Selection of cell populations with the intended genotype after genome editing, i.e. with CRISPR-Cas9. CellCelector Automated Cell Selection and Retrieval Platform is a perfect tool for targeted cell isolation and single cell cloning.
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Grow and expand cells in adherent or suspension cell cultures utilizing growth factors and cytokines to obtain sufficient supply for research and development needs. The Incucyte® Live-Cell Analysis System offers a label free, non-invasive approach to long-term monitoring of cell morphology and growth without removing cells from the incubator. Whether you are working with adherent or suspension cell lines, cultured cells need to be regularly passaged or sub-cultured. As all manipulation steps, passaging presents a contamination challenge. A good cell culture praxis includes testing for microbial contaminations on a regular basis.
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Advanced cell models using two-dimensional (2D) in vitro cell cultures usually utilize iPSC and MSC cultures. For iPSC generation adult somatic cells are used and genetically reprogrammed back into pluripotent state making them capable of becoming one of many types of cells inside a patient’s body.MSCs can be isolated from a variety of tissues, such as umbilical cord, endometrial polyps, bone marrow, adipose tissue, etc. For therapeutic development, i.e. CAR-T therapies the patient´s own (autologous) or donor´s T -cells are collected (allogenic).In most case gene editing is used for reprogramming of somatic cells or creating genetically modified i.e. knock-out, knock-in cell lines.
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3D cell models such as organoids and spheroids more closely recapitulate human cellular ecosystems which greatly improves clinical translation of new treatments.
Organoids are self-organizing, three-dimensional systems which retain many physiological characteristics of the native tissue from which they are derived. Accordingly, these miniaturized models have significant advantages over the use of traditional immortalized cell lines in providing accurate information on human disease modelling and can be used in the fields of drug screening, disease modeling, personalized medicine and many others.
Spheroids are simpler structures and less differentiated but still aggregation of one or more cell types in 3D cell cultures.
The goal of in vitro 3D cell culture is to enable more physiologically relevant and translational models in a wide range of biomedical applications including disease modeling, drug discovery and screening as well as personalized medicine. Through comprehensive characterization, scientists can develop a deeper understanding of biology, ensure more robust maintenance of stem cells during extended cultures and monitor cells at every step of the 3D organoid and spheroid workflow.
Evaluation of Critical Quality Attributes (CQAs) i.e. organoid/ spheroid size, sphericity, compactness, and viability is crucial to ensure the quality and reproducibility of the experiment, such us disease modeling, drug discovery and screening. Some parameters are tested throughout the culturing process including organoid health, counts and testing for microbial contaminations is recommended at several steps to ensure reliability and reproducibility of results. Sophisticated, quantitative analytical solutions which provide multi-parametric assays such Incucyte® Live-Cell Analysis and iQue®3 High-Throughput Cytometry platforms can greatly support the assessment of critical quality attributes.
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Creation of advanced cell-based assays for downstream analysis and specific applications for the research area of interest such us 3D disease modeling, drug discovery and compound screening.
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Further development and growth of organoids and spheroids enabling the formation of more complex structures. Specialty media, growth factors and cytokines support the expansion, differentiation, and maturation of 3D cell cultures. During extended cultures is important to monitor the formation and growth of organoid and spheroid cultures to maintain their health and desired characteristics.
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Selection and transfer of individual organoids and embedding organoids into 96- or 384-well plates. The CellCelector Automated Cell Selection and Retrieval Platform enables the automated scanning, detection and gating of complex 3D structures.
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Cells continue to differentiate and organize themselves into an organ-like structure in vitro. The Incucyte® Live-Cell Analysis System can be used to kinetically and objectively quantify multiple parameters to assess organoid growth and health during extended passaging.
The 3D cell culture process begins with seeding dissociated tissue-derived or stem cells into a 3D semi-solid extracellular matrix expanding these cells in defined media.
Characterization of organoid formation using real-time kinetic measurements on the Incucyte® Live-Cell Analysis System provides the ability to objectively assess the impact of growth conditions and seeding densities and define the conditions necessary to maximize expansion.
Pluripotent stem cells (PSCs), with their unique ability to differentiate and specialize into all the cell types of the body (referred to as pluripotency), are the most promising starting cell type source for building organoids. With appropriate signal stimuli in 3D in vitro culture systems, pluripotent stem cells can readily develop into organoids.Patient-derived cancer or healthy cells are increasingly used for organoid and spheroid generation due to their advantages compared with immortalized cell lines such us greater biological relevance. Gene editing technologies can be used to correct mutations in primary cells to study gene correction in patients.
Close the “bench to bedside” gap with innovative tools providing data and translational insights you can trust.
Reduce the risk of downstream failures when utilizing advanced cell models with our high-throughput instruments and automated solutions.
Explore leading solutions to accelerate progress from R&D, to Process Development and Manufacturing, safely and cost effectively.
Explore our portfolio of high-quality cell culture media, buffers, transfection reagents, growth factors, and cytokines for research applications.
Our extensive range of ready-to-use transfection reagents is suitable for research involving most mammalian primary cells and cell lines. These best-in-class transfection products enable the analysis of protein functions, research into signaling pathways, genome editing, and efficient RNA interference for the transfection of guide RNA and Cas9 expression.
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Cytokines and growth factors are key immune system signaling molecules that regulate immune responses, inflammation, cell growth, and differentiation. Their research has advanced our understanding of immune-related diseases and led to the development of targeted therapies and vaccines.
Sartorius produces high-quality, recombinant cytokines and growth factors for research purposes, free from animal-derived components. These rigorously tested products ensure reliable results, consistent supply, and meet the highest purity and efficacy standards, with ISO9001-certified quality management.
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Sartorius ready-to-use media are designed to support the growth and maintenance of a variety of cells and cell lines while meeting the highest quality standards. Each lot is manufactured under a strictly controlled process according to a Product Master Record for lot-to-lot consistency.
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The Incucyte® Live-Cell Analysis System enables real-time live-cell analysis directly inside your incubator. It provides quantitative, label-free in vitro approach for characterization and analysis of 2D and 3D cultures and cell models without ever having to remove cells from the incubator. The Incucyte® makes the process of acquiring, viewing, analyzing and sharing images of living cells easier than ever before. Incucyte® 21 CFR Part 11 Software Module offers comprehensive security and electronic record keeping functionality
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The CellCelector Flex Platform is a fully automated cell imaging and picking system developed for screening, selection and isolation of single cells, clusters, spheroids and organoids as well as single-cell clones and adherent colonies.
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The iQue® 3 High-Throughput Screening Cytometer is a high-throughput suspension cell and bead analysis platform for rapidly profiling of immune-cell phenotype and function in drug discovery and development workflows. Ideal for screens where cells are precious or limited in number, iQue® 3 is the fastest way to generate high-content data from small samples and the iQue® 21 CFR Part 11 Software Module ensures compliance for regulated laboratories at every stage.
Microfluidics acquisition capability analyses samples as small as 1uL minimum sample volume requirement in a 384-well format with zero dead volume. Cell detection occurs at rates of thousands of cells per second.
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Octet®️ Label-Free Bio Layer Interferometry (BLI) technology uses optical biosensors to measure protein-protein interactions in parallel, without the use of detection agents. This robust and fluidics-free approach enables fast, real-time characterization of expressed proteins, even in complex and unpurified samples.
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Testing cell cultures for microbial contamination on a regular basis ensures culture performance and parameters remain consistent and reproducible.
Microsart® Mycoplasma, Bacteria and Fungi qPCR kits offer rapid, reliable and easy-to-use solutions for microbial contamination control in compliance with international guidelines. Get results in just three hours!
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Avoid failures in your pre-clinical / clinical pipeline with translational research tools which recapitulate closer the human in vivo environment enabling better predictive outcomes.
Understand complex biology with greater clarity by utilizing leading-edge technology for the imaging and flow cytometry characterization of 2D immune and stem cells as well as complex, self-organizing 3D models.
Monitor continuously your 2D and 3D cell colonies, organoids and spheroids as they form and grow over time in controlled environment and capture quantitative information about their morphology, differentiation and health.
Reduce downstream risks with data you can trust – from the start.
In order to effectively use organoid models specific and reliable culture and analysis methods are required; learn how.
Exploring the next generation of antibody therapeutics, such as antibody–drug conjugates (ADCs), bispecific antibodies and nanobodies.
Learn about the impact of proper pipetting on 3D cell cultures, and which techniques can improve the reproducibility of various experiments.
Measure T-cell response in 3D models with characterization reagents and validated spheroid washing and dissociation protocols.
Organoids are defined by three key characteristics: self-organization, spatially restricted cell fate decisions, and ability to perform at least one function observed in in vivo organ tissue.
Spheroids are also 3D cell culture models, but they represent simple structures of typically the same cell types thus providing a versatile tool for translational research and disease modelling. There is a higher order self-assembly in organoids as opposed to spheroid cultures, and the former is more dependent on a matrix for its generation.
The application with techniques initially developed for analysis with 2D cultures and 3D culture production and specification are seen as main challenges. Still limited by the technologies used to characterize these complex cell models and may introduce variability due to lack of environmental control and inadequate 3D Cell Culture protocols that compromise organoid or spheroid cell health.
The CellCelector Flex platform enables gentle picking of a wide range of organoid sizes, ranging from 80 μm to 3.5 mm with exceptionally low (1 μL) injection volumes of surrounding media into either 100% hydrogel, liquid media or any other medium. It automatically measures morphology before and after the picking and transfer to prove that the organoids retain their morphology and structure due to the very gentle transfer method.
Real-time tracking of changes in organoid eccentricity (object roundness) and darkness (object brightness) with the Incucyte® Live-Cell Analysis System enables rapid, unbiased assessment of optimal culture passage periods.
The iQue® spheroid immune cell killing (ICK) workflow measures subset analysis and cytokines quantification to examine the activation, killing, exhaustion, and memory profiles of T cells during co-culture with single spheroids. The workflows utilize the iQue® Advanced High-Throughput Flow Cytometer with associated suite of T cell characterization reagent kits, and validated spheroid washing and dissociation protocols to provide an end-to-end solution.
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