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Cat. No.: 3DCCK-5 (for 5plts)

Description

3D Cell Culture Coating Kit for 96-Well Plate with Round Bottom is a convenient and efficient kit for preparing U-bottom 96-well plates for 3D cell culture, supporting the continuous formation and cultivation of cell spheroids. This kit is particularly suitable for providing a 3D growth microenvironment for various cells, allowing them to form cell spheroids from single-cell suspensions. These include tumor spheroids, tumor organoids, neurospheres, and leukocyte spheroids. The cell culture plates coated with this product can generate cell spheroids of relatively uniform size, making them widely applicable in tumor research, drug screening, organoid construction, and tissue engineering.

This kit includes a 3D cell culture coating solution and U-bottom 96-well plates for constructing a 3D cell culture environment. The main component of the 3D cell culture coating solution is a special high-molecular-weight polymer that serves as a non-adhesive matrix to coat the containers. It quickly forms a hydrophilic coating on the container surface, inhibiting cell adhesion to the growth surface of the container. When used with 96-well cell culture plates, this product ensures consistent and reproducible 3D cell growth, significantly reducing variability. The process for coating cell culture plates is simple and user-friendly, and the subsequent experimental procedures for cell culture are similar to traditional cell culture experiments.

Traditional cell culture predominantly takes place in a two-dimensional (2D) format. However, cells cultured in 2D exhibit significant differences in growth patterns, morphology, differentiation, and functionality compared to their true shapes and structures under physiological conditions in vivo. This may reduce the reliability of experimental results due to the lack of cell structure and tissue morphology. Three-dimensional (3D) cell culture can better mimic the in vivo microenvironment of cells, more accurately represent in vivo tissues, and more genuinely reflect cell-cell and cell-matrix interactions. Cell responses to exogenous and endogenous stimuli in 3D culture are closer to their reactions in vivo. Consequently, 3D cell culture becomes a more valuable and reliable in vitro experimental model, providing results more consistent with in vivo experiments.

3D tumor cell models are increasingly used to understand disease mechanisms and drug development. Tumor cells cultured in 2D can uniformly receive nutrients and oxygen on their single-sided membranes, while the inner cells of 3D cultured tumor cell clusters have fewer opportunities to access nutrients and oxygen, forming natural nutrient and oxygen gradients. This better simulates the in vivo microenvironment, making 3D cultured tumor spheroids or organoids more similar to in vivo tumors. This advantage supports small molecule drug screening and tumor-related molecular mechanism research, better predicting in vivo responses, efficacy, or toxicity of drug treatments. Additionally, 2D tumor cell models have limitations in vitro, as passaging may result in the loss of genetic heterogeneity and the selection of dominant clones, reducing clinical relevance. Compared to 2D cell models, 3D cell spheroids or organoids often provide more reliable research results, simplifying and accelerating drug evaluation processes. Since the first establishment of small intestinal organoids in 2009, 3D cell and organoid research has expanded to many tissue systems and become one of the most popular fields in life sciences.

Features

• This kit is simple and convenient to use. The kit includes high-quality, sterile SBS3D™ 96-well culture plates (round-bottom with lid, untreated surface, individually packaged), also known as U-bottom 96-well plates. With just one step, you can complete the construction of a 3D cell culture environment by coating the U-bottom 96-well plates with the 3D cell culture coating solution. The entire coating process takes a minimum of about 30 minutes. After coating, seed and culture the cells using conventional cell culture methods. Generally, you can obtain relatively tight cell spheroids within 24-48 hours of continued culture. The formation time of cell spheroids may vary for different types of cell culture plates. The size of the cell spheroids can be controlled by the initial seeding density of the cells.
• This kit has a wide range of applications and is suitable for the 3D culture of many common cells, with fast culture speed. The kit supports different cell types, including both adherent cells (e.g., HCT-116, HeLa) and suspension cells (e.g., K562, MOLM-13). Cells seeded in the U-bottom 96-well plates coated with this kit can show significant aggregation within 2 hours, and the spheroids become more compact over time. After 48 hours of culture, a relatively tight cell spheroid can be formed, and the size of the spheroid is proportional to the number of seeded cells. Compared to flat-bottom multi-well plates or culture dishes, which generally require more than 3-5 days to form cell spheroids, the U-bottom structure is more efficient.

Components

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Storage

Store at 4ºC, the product is effective for one year, while store at room temperature, the product is effective for six months. The 3D cell culture coating solution can be stored at -20ºC for a longer duration. The U-bottom 96-well plates are effective for three years when stored at room temperature.

Precautions

• If the 3D cell culture coating solution is stored at -20ºC, it must be brought to room temperature before use.
• The 3D cell culture coating solution contains a certain concentration of ethanol, which is volatile. It must be sealed during use and storage.
• When adding the 3D cell culture coating solution to the U-bottom 96-well plates, avoid forming bubbles. If bubbles do appear, carefully puncture them with a pipette tip, as they may affect the coating efficiency of the U-bottom 96-well plates and, consequently, the cell culture results.
• For immediate coating, the standing time after adding and removing the coating solution should be no less than 10 minutes, and extending the time is preferable. Otherwise, it may affect the culture results.
• The spheroid formation speed, compactness, and morphology of 3D cells are closely related to cell type and cell number. Different cell types and cell numbers may result in significant differences in spheroid formation speed and state.
• Although this product can be used for immediate coating, for better culture results, it is recommended to coat the plates in advance and dry them in a drying oven. If the dried plates cannot be used immediately, seal them and store them at 4ºC for up to 3 months; at room temperature for up to one week; and at 37ºC for up to one day.
• This product is intended for scientific research use by professionals only and is not for clinical diagnosis or treatment, food, or drugs. Do not store in residential areas.
• For your safety and health, wear a lab coat and disposable gloves while handling.