Prostate Organoid Culture in Microbeads

Advances in three-dimensional (3D) tissue organization models

The past decade has brought significant advances in prostate cancer (PCa) research. With the increased understanding of the origin and the molecular landscape of PCa, there has been an encouraging trend of precision medicine-based approaches to treat advanced PCa. 

As we already know, tissues and organs are multicellular structures that self-organize in three dimensions (3D). Cells within a tissue, such as glandular tissue, interact with neighboring cells and the extracellular matrix (ECM) through biochemical and mechanical signals that maintain the specificity and homeostasis of biological tissues. 

While traditional 2D cultures on rigid surfaces fail to reproduce cellular behavior in-vivo, 3D matrices are becoming increasingly popular supports for cell culture because they mimic the complex environment that supports the physiological functions of cells to better predict in-vivo responses, thus limiting the need for animal models. 

Organoids as Models for Prostate Cancer Research 

Organoids are relevant models to mimic the complex in-vivo environment that supports cell physiological and pathological behaviors. For instance, 3D epithelial organoids recapitulate numerous features of glandular tissues including the development of fully differentiated acini that maintain apicobasal polarity with the hollow lumen. Therefore, researchers have been focusing on the production of prostate organoid culture to better understand the complexity of PCa initiation and progression.  

Effective genetic engineering in prostate organoid culture would provide new insights into organogenesis and carcinogenesis, helping us to decipher the key genetic networks underlying epithelial differentiation and polarity, and allowing us to better understand how they may be altered in pathological states such as cancer. 

Challenges in 3D Transfection of Organoids 

However, direct 3D transfection on already-formed organoids remains challenging. One limitation is that organoids are embedded in the extracellular matrix and grow into compact structures that hinder transfection using traditional techniques. To address this issue, Laperrousaz, B. et al. (2018) have developed an innovative approach for transgene expression in 3D prostate organoid culture by combining single-cell encapsulation in Matrigel microbeads using a Fluigent microfluidic device and electroporation.  

Laperrousaz, B. et al. (2018) demonstrated that direct electroporation of encapsulated prostate organoid cultures reach up to 80% of transfection efficiency when combining Fluigent’s technology for organoid generation and efficient 3D transfection. They were also able to validate the role of p63 and PTEN as key genes in acinar development in breast and prostate tissues confirming that this encapsulation and transfection method opens up new perspectives for flow-based high-throughput genetic screening and functional genomic applications.