CellHD-256 Hanging Drop Chip for 3D Spheroid Culture

The formation of three-dimensional (3D) cellular spheroids is one of the most straightforward methods to create in vivo-like cell culture-based assay in biomedical research. The 3D spheroids have a concentric circle structure and closely mimic the microphysiology in vivo, through which scaffold-free cultured 3D cells with desired cell density can be obtained. Common methods to produce cell aggregates include stirring suspension, magnetic suspension, hanging drop culture, etc., which are limited by the variation in spheroids size and cell number, the high labor intensity, the high shear force, and the difficulty of massive production. Among various 3D sphere generation techniques, hanging drop culture has been widely adopted due to its single spheroid formation and high reproducibility. The main obstacle for hanging drop method to culture 3D cell spheroids is the culture tool, and using conventional dishes makes media exchange difficult. Microfabrication based tools for hanging drop have been developed with the characteristic of forming a large number of aggregates with uniform size, less laborious, easy control, and amenable to high-throughput screening.

CellHD-256 Hanging Drop Chip

CellHD-256 Hanging Drop Chip

Amerigo Scientific offers CellHD-256 Hanging Drop Chip, a user-friendly 3D cell culture platform designed to provide life science researchers and drug developers with an efficient and convenient solution for 3D spheroid generation and cultivation. CellHD-256 is a chip-based microfluidic technology for high-throughput and uniform production of 3D cell spheroids. It provides a high throughput-256 culture drops for cellular spheroids simultaneously on a single chip, and less medium is used during cultivation.

Principle of CellHD-256

CellHD-256 uses the principle of hydraulic differential to form hanging drops. Due to gravity and the inherent adhesion properties of cells, the hanging drop technique can cause cells to spontaneously aggregate into spheroids within droplets. Open holes in the bottom of the microchannel in a chip is utilized to form medium drops, and suspended cells can aggregate at the bottom of each drop. The concave shape of the droplets is necessary to form a single-cell sphere in each droplet and is controlled by the height of the medium in the medium reservoir. The higher the liquid level in the reservoir, the greater the curvature of the droplet. It is recommended to fill the reservoir with medium close to the top of the reservoir (approximately 1 mm from the top).

The cell suspension concentration determines the formation of the initial cell spheroid size, so the concentration should be determined according to the cell type and experimental design. We recommend a cell suspension concentration ranging of 1~10 × 10⁴ cells /mL for seeding.

Features of CellHD-256

CellHD-256 utilizes a patented microchannel design for the high-throughput generation of hanging drops in which uniform cell aggregates can be formed and grow into 3D cell spheroids on the chip.

Applications of CellHD-256

• Drug Discovery and Screening

CellHD-256 hanging drop chips can be used for high-throughput drug screening, accelerating the drug discovery process and improving the efficiency and success rate of drug discovery.

• Cancer Research

The cell spheroids cultured by CellHD-256 can simulate the 3D growth environment of tumor cells and be used to study the mechanism of tumor invasion, metastasis, and response to anticancer drugs.

• Stem Cells and Tissue Engineering

CellHD-256 supports the 3D culture and differentiation of stem cells, facilitating the development of novel strategies for tissue repair and regenerative medicine.

• Cell Interaction Studies

CellHD-256 can be used to study the interaction of different cell types in a 3D environment, such as between immune cells and tumor cells.

Advantages of Hanging Drop Method for 3D Spheroid Culture

Hanging drop method is a widely used technique for generating 3D cell spheroids. Compared to other 3D culture methods, this approach offers distinct advantages.

• Uniform and controllable spheroid size: Through precisely controlling the number of cells in each hanging drop, spheroids of consistent size can be generated, ensuring the reproducibility and reliability.
• No requirement for scaffolds or matrices: Due to gravity, cells can concentrate at the bottom of the drop and form a spheroid, thus simplifying the experimental workflow without additional scaffold materials using the hanging drop method.
• Avoidance of exogenous material interference: Since no external scaffolds or matrices are required, potential interference from these materials on cell behavior is minimized. This ensures more accurate and cell-specific experimental data.
• Simple operation and low cost: The method requires no complex equipment or expensive consumables. Basic cell culture materials are sufficient, making it cost-effective and accessible for laboratories.
• Compatibility with diverse cell types: The hanging drop method is widely applicable to various cell types, such as primary cells, tumor cells, and stem cells.
• Effective simulation of 3D microenvironment: Spheroids formed via hanging drop culture could mimic the in vivo 3D cellular microenvironment. The spatial organization and interactions within spheroids resemble in vivo physiological conditions, enabling studies on cell function, signaling, and drug responses.
• Scalability and suitability for high-throughput screening: The method allows simultaneous processing of numerous samples, making it ideal for high-throughput applications such as target discovery and drug screening.
• Ease of observation and analysis: Spheroids suspended in droplets are readily observable under microscopes, facilitating real-time monitoring of formation and growth. Sampling and downstream analyses are also simplified.