Protocol for production of homogeneous iPSC spheroids and microtissues using the suction technique

Area of Science:

• Biotechnology
• Stem Cell Research
• Drug Discovery

Background:

• Three-dimensional (3D) cell cultures offer more physiologically relevant models compared to traditional 2D cultures.
• 3D cultures are increasingly vital in drug discovery for improved predictive accuracy.
• Induced pluripotent stem cells (iPSCs) are valuable for generating patient-specific cell models.

Purpose of the Study:

• To introduce a reproducible protocol for generating homogeneous induced pluripotent stem cell (iPSC) spheroids and microtissues.
• To detail the suction technique for efficient cell sedimentation and spheroid self-assembly.
• To provide a method for evaluating the quality of the generated spheroids.

Main Methods:

• Development and preparation of a specialized suction device.
• Optimization of cell preparation and seeding for uniform distribution.
• Application of the suction sedimentation technique for cell aggregation.
• Step-by-step guidance on spheroid self-assembly and subsequent evaluation.

Main Results:

• Successful generation of homogeneous iPSC spheroids and microtissues.
• Demonstration of the suction technique's efficacy in producing consistent 3D cell aggregates.
• Establishment of a reliable protocol for 3D cell culture in drug discovery.

Conclusions:

• The suction technique provides an efficient and reproducible method for creating iPSC spheroids.
• This protocol facilitates the development of advanced 3D cell models for drug discovery.
• The presented method contributes to the advancement of stem cell-based research and therapeutic development.