Organoid Research

Stevan Hoyle

Feb 19, 2018

Cell Culture

Author: Stevan Hoyle

Stevan Hoyle

Introduction

It has long been known that cells cultured in two dimensions (2D) are not representative of the in vivo situation due to their growth being restricted to a flat surface. Furthermore, 2D structure minimises cell-cell and cell-matrix interactions. This cause the distinctive phenotype to be lost and the cells to react differently to external cues. However, 2D cell culture remains one of the mainstays of research programmes worldwide. Because numerous failures in translating effective therapies from cell models into humans can partly be attributed to a reliance on 2D cell culture, a more physiologically relevant human models is developed. This leads to the establishment of three-dimensional (3D) cell culture methods during the past few decades. This 3D cell culture, have afforded significant advancement of a wide range of therapeutic strategies and a key breakthrough within this field has been the development of organoid technology.

Organoid technology

The term organoid is used to describe a 3D cell culture which consists of organ-specific cell types. These simplified mini organs share the in vivo architecture and functionality of the original tissue, making them ideal model systems for research. Organoids can be generated from primary tissue samples which contain adult stem cells, from embryonic stem cells (ESC) or from induced pluripotent stem cells (iPSC), and their development represents one of the most exciting recent advancements in stem cell research. Additionally, the number of publications referencing organoids has increased dramatically in the last ten years, with their use described for disease modelling, drug screening, regenerative therapy, host-microbe interactions and for studies of organogenesis and morphogenesis.

Patient-derived organoids hold huge potential for increasing our understanding of various disease states and predicting treatment response. They also open up many possibilities for personalised medicine. For example, transcriptional and proteomic analyses performed on organoid models derived from normal and neoplastic pancreas tissues have been used to identify key genes associated with progression of pancreatic cancer1, while our understanding of the neurobiology of Zika virus has advanced through the use of brain organoids; organoid modelling followed by ZIKV infection has suggested neural progenitor cells to be particularly susceptible to infection2. In another study, the CRISPR/Cas9 genome editing system has been used to correct the CFTR locus by homologous recombination in cultured intestinal stem cells of cystic fibrosis patients, providing proof of concept for gene correction in primary adult stem cells derived from patients with a single-gene hereditary defect3.

Organoid research

As organoid research has increased in popularity, the range of supporting tools and reagents has grown accordingly. A number of companies offer high quality media products, matrix components, growth factors and small molecules designed specifically for organoid generation, while others provide protocols and technical support to promote the successful establishment of organoid cultures. We should of course remember that organoid culture, although extremely sophisticated, is in reality simply an advanced form of cell culture; as such many existing tried and trusted reagents are applicable to this research field. For example, PeproTech’s extensive range of cytokines and growth factors may be used in organoid research, and their epidermal growth factor (EGF), Noggin and R-Spondin proteins were all cited in a recent publication describing the establishment of different 3D systems, including organoid culture, to culture gastrointestinal epithelium.

References

  • Boj S. F. et al. Organoid models of human and mouse ductal pancreatic cancer. Cell. 2015;160(1-2):324-338. doi:10.1016/j.cell.2014.12.021
  • Li H. et al. The Neurobiology of Zika Virus. Neuron. 2016;92(5):949-958. doi:10.1016/j.neuron.2016.11.031
  • Schwank G. et al. Functional repair of CFTR by CRISPR/Cas9 in intestinal stem cell organoids of cystic fibrosis patients. Cell Stem Cell. 2013;13(6):653-658. doi:10.1016/j.stem.2013.11.002
  • Pastuła A. et al. Three-Dimensional Gastrointestinal Organoid Culture in Combination with Nerves or Fibroblasts: A Method to Characterize the Gastrointestinal Stem Cell Niche. Stem Cells Int. 2016;2016:3710836. doi:10.1155/2016/3710836

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