Cell Counting Kit-3D
$248.00 - $868.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: CCK3D-5 (for 5ml)
Cat. No.: CCK3D-25 (for 25ml)
Description
Cell Counting Kit-3D, abbreviated as CCK-3D assay kit or CCK3D assay kit, is a rapid and highly sensitive detection kit based on the WST series chromogenic substrate for assessing 3D cell viability, proliferation, and cytotoxicity. This assay kit is suitable for measuring absorbance at around 450nm and typically takes about 1 hour to complete the testing.
Traditional cell cultures are primarily conducted in two dimensions (Two-Dimensional, 2D). However, as research has advanced, scientists have discovered significant differences in the growth pattern, morphology, cell structure, differentiation, and biological functions of cells cultured in 2D compared to cells in their natural physiological conditions inside the body. These differences can lead to substantial discrepancies between experimental results and the actual in vivo situation. Three-dimensional (Three-Dimensional, 3D) cell culture techniques provide a better simulation of the microenvironment in which cells thrive in the body. Cells cultured in 3D can maintain their normal biological functions, better mimic in vivo tissues, and more accurately represent cell-cell and cell-matrix interactions. Their responses to external and internal stimuli are also closer to what occurs in the body. As a result, 3D cell culture has become a superior in vitro model for cell experiments, yielding experimental results that are closer to or consistent with in vivo experiments.
3D tumor cell models are increasingly used to understand disease mechanisms and drug development. In 2D cell cultures, tumor cells receive nutrients and oxygen uniformly from one side of the cell membrane. In contrast, 3D-cultured tumor cell clusters have fewer internal cells that receive nutrients and oxygen, creating a natural gradient of nutrients and oxygen. This better simulates in vivo microenvironment conditions. Therefore, 3D culture systems such as tumor spheroids or organoids are more effective at simulating in vivo tumors. They are better suited for small molecule drug screening and the study of tumor-related molecular mechanisms. Additionally, they can accurately predict in vivo responses, efficacy, or toxicity of drug treatments. In comparison to 2D cell models, 3D cell spheroids or organoids often provide more reliable research results, simplifying and expediting the drug evaluation process. Since the establishment of the first small intestine organoids in 2009, research on 3D cells and organoids has expanded to encompass many tissue systems and has become one of the hottest fields in life sciences.
As 3D cell culture techniques have matured, they have become widely adopted research tools and are increasingly popular. They find applications in various fields, including basic research and drug discovery, and the assessment of cell viability under 3D culture conditions has become a significant challenge.
The WST series chromogenic substrates, such as WST-1 and WST-8, are a group of compounds similar to MTT. In the presence of electron-coupling reagents, they can be reduced by certain dehydrogenases inside the mitochondria to produce orange-yellow formazan. The color of the formazan is darker as cell proliferation increases and lighter as cell toxicity increases. For the same type of cells, the color intensity is linearly related to the number of cells. This assay kit has been optimized in various aspects, and the absorbance measurement of formazan allows for highly efficient, linear, and accurate assessment of overall cell viability in 3D cell spheroids.
This assay kit can be used for detecting cell proliferation induced by factors like cytokines, as well as for assessing cytotoxicity induced by anti-cancer drugs or other toxic agents that inhibit cell growth induced by certain drugs.
Features
- This assay kit offers a very convenient detection process. The kit contains a pre-prepared CCK-3D solution in a single tube, requiring no additional preparation steps. There is no need for the use of isotopes, and all the detection steps can be completed within the same 96-well plate. Cell washing, cell collection, or additional steps to dissolve formazan are not necessary. It can be used for high-throughput screening of large sample batches.
- Phenol red and serum have no significant impact on the assay results of this kit.
- This product exhibits minimal toxicity to cells. After adding the CCK-3D solution and color development, readings can be taken repeatedly at different time intervals using a plate reader, offering flexibility in determining the optimal assay time.
- This product is suitable not only for the analysis of small sample quantities but also for high-throughput screening of large sample batches. It can be used with 3D cell spheroids cultured using various methods, including cell suspension culture plates, ultra-low attachment cell culture plates, flat plates coated with extracellular matrix or Matrigel, and plates coated with agarose, among others.
Notes
- Repeated freeze-thaw cycles may reduce the effectiveness of this assay kit. Although it has been tested that freezing and thawing the kit up to three times does not significantly affect its performance, for optimal results, it is advisable to aliquot the kit after the initial thawing. During repeated freeze-thaw cycles, a slight precipitation may occur, in which case it should be equilibrated to room temperature and dissolved as much as possible.
- Temperature fluctuations can affect signal intensity and signal stability. Both the cells and CCK-3D solution should be equilibrated to room temperature before the assay. CCK-3D solution can be thawed and mixed at room temperature not exceeding 25°C or in a water bath at 20-25°C before use.
- As the assay is conducted in a 96-well plate, it's important to be aware of potential evaporation issues, especially if the cell culture duration is extended. To mitigate this, you can either discard the wells around the edges of the 96-well plate, as those are most prone to evaporation, and replace them with an equal volume of PBS, water, or culture medium. Alternatively, you can place the 96-well plate near a water source within the incubator to reduce evaporation.
- 3D cells typically form spheroids in the wells of the 96-well plate. The orange-yellow formazan generated after adding the CCK-3D solution tends to aggregate on and around the cell spheroids. Before measurement, gently tap the 96-well plate to ensure even distribution of formazan in the cell culture medium. Otherwise, the signal values obtained may be biased, affecting the accuracy of the results.
- This assay kit relies on a reaction catalyzed by dehydrogenases, so reducing agents (such as some antioxidants) can interfere with the assay. If there are significant levels of reducing agents in the test system, they should be removed.
- For the assay, standard transparent 96-well or 384-well plates are sufficient. You can also use ultra-low attachment culture dishes or multiwell plates designed for 3D cell culture, and transfer the 3D cells to standard transparent multiwell plates for measurement. Various ultra-low attachment culture dishes and multiwell plates suitable for 3D culture are available, please refer to related products.
- Before using a plate reader for the assay, ensure there are no air bubbles in each well, as they can interfere with the measurement.
- This product is intended for scientific research use by trained professionals only and should not be used for clinical diagnosis or treatment, nor for food or drug purposes. It should not be stored in residential areas.
- For your safety and health, please wear appropriate laboratory attire and disposable gloves when handling this product.
Storage
Store in a light-protected manner at -20°C. Valid for one year.
Related:
- Cell Counting Kit-8 (CCK-8)
- Enhanced Cell Counting Kit-8
- Super-Enhanced Cell Counting Kit-8
- Cell Counting Kit-3D
Only for research and not intended for treatment of humans or animals
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