Instant Cells Assay Kits
Creative Biolabs has the assays you can rely on for high throughput screening, lead optimization, characterizing and discovering targets, and uncovering the complexity of disease pathways. We can offer membrane protein in vitro assay kits that save valuable laboratory time and is ideal for high throughput screening.
Background of Membrane protein
Common proteins that are a component of or interact with biological membranes are known as membrane proteins. About one-third of all human proteins are membrane proteins, and more than half of all medications target them. Membrane proteins are widespread and crucial in medicine. However, compared to other protein classes, figuring out membrane protein structures is still difficult, largely because it is challenging to create experimental circumstances that can maintain the protein's right conformation when it is separated from its natural environment.
Features of Instant Cells Assay Kits
A collection of recombinant cell lines encoding pharmaceutically important receptors have been created by Creative Biolabs. The targets are frequently employed for lead identification, selectivity testing, and small molecule compound screening. The cell lines don't need to be cultured before use; they can be utilized right away for automated and manual patch clamp setups after thawing.
Published Data
| Paper Title | Assay-ready Cryopreserved Cell Monolayers Enabled by Macromolecular Cryoprotectants |
| Journal | Biomacromolecules |
| Published | 2022 |
| Abstract | Cell monolayers facilitate the discovery and screening of new drugs and the study of cell biology and disease at a fundamental level. However, the current state of cryopreservation technology does not permit cells to be frozen while attached to tissue culture plastic. For the desired application, cells must be thawed from suspension, cultured for several days or weeks, and then transferred to multiwell plates. This inefficient procedure consumes a significant amount of time that could be spent conducting biomedical research or other value-adding tasks. In this report, researchers demonstrate that a synthetic macromolecular cryoprotectant enables the routine, reproducible, and robust cryopreservation of biomedically significant cell monolayers in tissue culture multiwell plates that adhere to industry standards. The cells are simply thawed in media and placed in an incubator, where they will be ready for use within 24 hours. Post-thaw cell recovery values for three cell lines were greater than 80%, with minimal variation between wells. The cryopreserved cells maintained healthy morphology, membrane integrity, proliferative capacity, and metabolic activity; exhibited marginal increases in apoptotic cells; and responded favorably to doxorubicin-based toxicological stress. These findings confirm that cells are "test-ready" 24 hours after thawing. Overall, they demonstrate that macromolecular cryoprotectants can solve a long-standing cryobiological problem and have the potential to transform routine cell culture for biomedical research. |
| Result |
Over 75% of cells were recovered 24h post-thaw, regardless of cell seeding density (12.5k–100k cells per well). Cryopreservation studies focus on confluent cell monolayers because of higher recovery values. For cell viability assays, a linear response between seeding density and signal output (usually absorbance or fluorescence) is required. Cryopreserving cells at multiple seeding densities is crucial for assay development and toxicological screening. A resazurin reduction metabolic assay was performed on nonfrozen and frozen A549 and HepG2 cells at varying seeding densities to determine the quantity of cells required to freeze to ensure a linear output with resazurin reduction to resorufin and allow its use for drug screening. Both cell lines metabolized resazurin to resorufin 24h post-thaw and had a linear relationship between cell density and absorbance/fluorescence. A549 metabolic activity was unaffected by freezing, but HepG2 was halved 24h post-thaw. Post-thaw, osteoblasts (MG-63), ovarian tissue, and mesenchymal stem cells reduce metabolic activity by half or more. Both A549 and HepG2 cells remained functional for drug screening cell viability assays.
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Fig.1 Assay optimization and metabolic activity of cryopreserved cells.1,2
References
- Tomás, Ruben MF, et al. "Assay-ready cryopreserved cell monolayers enabled by macromolecular cryoprotectants." Biomacromolecules 23.9 (2022): 3948-3959.
- Image retrieved from Figure 3 " Assay optimization and metabolic activity of cryopreserved cells. " Tomás, et al. 2022, used under CC BY 4.0. The original image was modified by extracting and the title was changed to "Assay optimization and metabolic activity of cryopreserved cells.".
