Organoid Research Techniques: Evolution and Applications
Interested in learning more about organoids? Download this practical guide on organoid research techniques.
This e-book covers the evolution of organoid techniques, from their original development and description to updated applications and future outlooks. Discussion of key milestones and annotated reading lists provide further background on many of the topics discussed, as well as other areas for the reader to continue exploring. Both adult stem cell-derived (ASC-derived) and pluripotent stem cell-derived (PSC-derived) organoids from various different tissues are discussed in detail, giving a complete overview of organoid technologies and some of their applications.
In this guide you'll find:
- An overview of the development of organoid technologies.
- Discussion of historic milestones and publications that led to the current state of organoid technologies.
- Detailed case studies on the application of both ASC- and PSC-derived organoid cultures.
- Discussion of challenges still facing organoid technologies.
- A look towards the future of organoid technologies and where they are headed.
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Pages 3-4, Introduction
The methods for culturing organoids are tissue鈥恉ependent, but some overall principles apply. A scientist embeds the starting material鈥攕uch as progenitor cells derived from pluripotent stem cells (PSCs) or adult stem cells (ASCs) harvested from tissue samples鈥攊n an extracellular matrix. The cells are maintained in culture media containing the nutrients and growth factors necessary to mimic the in vivo cellular environment. Under these conditions, the starting cells expand and self鈥恛rganize to build a 3D structure鈥攖he organoids鈥攚hich can be maintained for long periods of time. For some epithelial organoids (e.g., intestinal organoids), the cultures can be maintained indefinitely through routine passaging (i.e., breaking the organoids into small sections and reseeding them in new cultures).
Developing culture conditions for organoids is not a trivial matter. Certain conditions must be met for the organ鈥恟esident stem cells to be maintained and differentiated into the appropriate complement of cells. This fundamental property of organoid generation makes the system ideal for understanding organ development and the biology of the adult鈥 or tissue鈥恟esident stem cells that maintain a given tissue throughout life. Organoids also lend themselves to many other applications, such as cell biology, drug development, and the understanding of disease. Their wide range of applications, coupled with their potential to significantly reduce the use of animal models while allowing for facile experimentation on human cells, makes organoids valuable model systems. Consequently, organoid model systems are seeing rapid adoption in laboratories world鈥恮ide.
Figure 1. Light microscope visualization of a mouse intestinal organoid.
Source: 海角破解版 Technologies
Pages 12-13, In Practice
Organoid Types and Culture Conditions
There are two primary classifications of organoid systems: organoids cultured from PSCs and organoids cultured from adult stem or progenitor cells. PSC鈥恉erived organoids are generated by differentiation of PSCs to lineage鈥恠pecific progenitor cells that form the desired organoid. Organoid formation is then achieved by culturing these progenitors in conditions that mimic the in vivo developmental conditions of that organ. For brain organoids, by omitting or adding patterning factors, the organoid can either be allowed to self鈥恛rganize into an organoid with several different brain regions (e.g., cerebral organoids) or can be directed to form an organoid with specific brain regions (e.g., forebrain or midbrain organoids). The advantage of PSC鈥恉erived organoids is their easy鈥恡o鈥恆ccess starting material, because previously鈥恊stablished or patient鈥恠pecific PSC lines can be used. These organoids tend to model the developing, rather than the adult organ. ASC鈥恉erived organoids are cultured from tissue鈥恠pecific stem or progenitor cells that are responsible for maintenance of a given organ in vivo. The organoids grown from these progenitors tend to more closely model adult tissue and have the advantage that they can recapitulate both congenital and non鈥恈ongenital disease states, including modeling epigenetic and tumor signatures.
Both classes of organoids are cultured in specific cell鈥恈ulture media within an extracellular matrix that is required to support the 3D structure. The cell culture medium used depends on the type and tissue of the organoid, and in general mimics the signaling environment the cultured cells would experience in vivo. For PSC鈥恉erived cerebral organoids, for example, the culture conditions mimic the signaling present in the developing brain, allowing the neural progenitors present in early stages of the organoid to differentiate and self鈥恛rganize into the laminar structure of the cerebrum. In similar fashion, the culture conditions for ASC鈥恉erived intestinal organoids mimic the signaling present at the intestinal crypt base, the stem cell niche of intestinal stem cells. This allows for self鈥恟enewal of the stem cell population as well as differentiation of a subset of this population to form the cellular complement of the intestine.
Page 14, In Practice
Figure 2. Adult stem cell-derived organoids: (a) Human colonic organoids; (b) mouse prostate organoids; (c) pancreatic exocrine organoids; and (d) hepatic progenitor organoids.
Source: 海角破解版 Technologies
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