3D Neural Spheroids Culture Technology

Creative Biolabs is always the leader in 3D neural spheroids culture services and offers a variety of technologies to help you grow 3D neural spheroids quickly and efficiently. In addition, we also provide functional tests and a one-stop service for your research and development to bring more physiological relevance.

3D Neural Spheroids

Since the growth patterns and functions of neural stem cells (NSCs) under traditional 2D culture conditions are different from those in vivo, finding effective therapeutics for neurological diseases remains challenging. Advanced 3D cell culture techniques have been widely adopted to mimic realistic physiological microenvironments, bridging the gap between in vitro and in vivo experimental results.

Due to the possibility of contact differentiation in cell adherence culture, an undifferentiated state is a primary prerequisite for studying the characteristics of stem cells. Therefore, neural spheroids are considered as a suitable in vitro model for studying NSCs. Neural spheroids are multi-cellular aggregates of NSCs that spontaneously self-assemble or proliferate in vitro without scaffolds. 3D neural spheroids are similar to natural tissues both in morphology and function, so as to better simulate the growth of cells in vivo, more intuitively reflect the biological functions of NSCs, and more accurately construct the target neural tissue model. 3D neural spheroids can be formed by 3D suspension drop culture and ultra-low adhesion culture.

• 3D Suspension Drop Culture

When a drop of cell suspension is added above the hole, the geometry of the 3D droplet culture plate guides the cell suspension through a small hole, resulting in the cell suspension becoming a droplet held by surface tension. This method requires a small amount of cell suspension (usually 20μL). The density of cells to be inoculated depends on the size of the neural spheroids required. Neural spheroids formation in the 3D suspension culture plate is divided into three processes: cell suspension distribution, suspension droplet formation, NSCs generate endogenous extracellular matrix and continuously aggregate to form neural spheroids. The drop inlet is located above each culture hole and can be used to change the medium and add exogenous extracellular matrix, growth factors, and small molecules.

Fig.1 Sequential stages of hanging-drop spheroid formation: (A) Cell suspension deposition, (B) Gravity-enforced cell aggregation, (C) Mature 3D spheroid assembly.^1,3

• Ultra-low Adhesion Culture

Ultra-low adhesion culture plane is a hydrophilic and uncharged plane covalently coupled with hydrogel to minimize cell attachment, protein absorption, and enzyme activity, and maintain the growth of NSCs in a suspended and non-adherent state. A single 3D neural spheroid of uniform and repeatable size can be generated by applying the package to each hole of the spheroid microplate. Neural spheroid can be easily analyzed by fluorescence staining, which is suitable for high-throughput screening and functional assay

Fig.2 Schematic of a Transwell co-culture system for evaluating cytotoxic drug permeability across an in vitro blood-brain barrier (BBB) model (MDCKII/MDR1 monolayer) into glioblastoma spheroids (LN-229), followed by 3D viability assessment.^2,3

Creative Biolabs can provide you with personalized 3D neural spheroids culture solutions to meet your experimental needs and purposes. Please contact us to discuss your project, and our professional team will provide you with the most professional laboratory services according to your needs.

References

1. Białkowska, Kamila et al. “Spheroids as a Type of Three-Dimensional Cell Cultures-Examples of Methods of Preparation and the Most Important Application.” Int J Mol Sci. 2020;21(17):6225.
2. Sherman, Hilary, and Ann E Rossi. “A Novel Three-Dimensional Glioma Blood-Brain Barrier Model for High-Throughput Testing of Tumoricidal Capability.” Front Oncol. 2019;9:351.
3. Distributed under Open Access License CC BY 4.0 without modification.