Dissecting the stem cell niche with organoid models: an engineering-based approach - PubMed (original) (raw)

Review

Dissecting the stem cell niche with organoid models: an engineering-based approach

Lyndsay M Murrow et al. Development. 2017.

Abstract

For many tissues, single resident stem cells grown in vitro under appropriate three-dimensional conditions can produce outgrowths known as organoids. These tissues recapitulate much of the cell composition and architecture of the in vivo organ from which they derive, including the formation of a stem cell niche. This has facilitated the systematic experimental manipulation and single-cell, high-throughput imaging of stem cells within their respective niches. Furthermore, emerging technologies now make it possible to engineer organoids from purified cellular and extracellular components to directly model and test stem cell-niche interactions. In this Review, we discuss how organoids have been used to identify and characterize stem cell-niche interactions and uncover new niche components, focusing on three adult-derived organoid systems. We also describe new approaches to reconstitute organoids from purified cellular components, and discuss how this technology can help to address fundamental questions about the adult stem cell niche.

Keywords: Cell culture; Engineer; Model; Niche; Organoid; Stem.

© 2017. Published by The Company of Biologists Ltd.

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Conflict of interest statement

Competing interests

The authors declare no competing or financial interests.

Figures

Fig. 1.

Fig. 1.

Advantages of organoid models for studying adult stem cells. (A) Organoids grown clonally from single cells can be used to prospectively identify adult stem cell populations based on the capacity of a cell to form organoids. (B) Organoids can be derived from human cells as well as non-human cells such as mouse or zebrafish, which allows modeling of human-specific stem cell biology and the identification of differences between human and non-human tissues. (C) In vitro culture allows in-depth experimental perturbation and imaging of stem cells in their surrounding niche. Different approaches include tightly controlled chemical or genetic manipulation, 3D imaging of live tissues over time (4D imaging), high-throughput combinatorial screening, and single-cell resolution imaging to analyze specific cell-cell interactions.

Fig. 2.

Fig. 2.

Advantages of engineered organoids for studying the stem cell niche. (A) Constructing organoids from purified cellular components allows the direct measurement of input cell properties and labeling of different input populations. (B) In addition to the advantages of classical organoid models (outlined in Fig. 1), controlled organoid engineering provides tight experimental control over the numbers and types of cells in the resulting tissue. (C) Engineered organoids can incorporate non-cellular material, such as sensors that dynamically measure properties such as mechanical forces and signaling pathway activation within live tissues.

Fig. 3.

Fig. 3.

Technologies to reconstitute organoids from purified cell populations. (A) Microwells. Cells can be centrifuged, flowed or injected into arrays of microwells to produce organoids that conform to the size and shape of the microwell. (B) Microfluidics. Individual cells or ECM components can be captured in aqueous droplets and combined to produce precisely sized spheroids that are amenable to high-throughput recovery and analysis. (C) 3D bioprinting. Cells suspensions or ECM components can be used as a printable ‘ink’, with control in the x, y plane over individual components. Multiple ‘inks’ can be loaded into the printer to create complex, patterned tissues. (D) Chemically programmed assembly. Cell surfaces can be chemically modified with single-stranded DNA or other bio-orthogonal molecules to program adhesion to surfaces or other cells, independent of endogenous cellular machinery. This technique can achieve single-cell resolution to create organoids with precisely controlled cell-cell interactions. (E) Engineered ECMs. Polymer hydrogels such as PEG can be tuned over a range of stiffnesses and topologies by varying monomer concentration, molecular weight and degree of crosslinking. Cell adhesion ligands (e.g. RGD integrin-recognition sequences) or proteolytically degradable sequences can be engineered back into the system.

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