Intestinal crypt organoids as experimental models

When it comes to studying the effect of food bioactives on gut health, one of the essential steps that needs to be assessed is characterizing specific effects of the bioactives on the physical barrier of the lumen, the gastrointestinal tissue. In addition to studying the effects on transport function (e.g. by using Ussing chambers or cell culture systems), it is of great interest to evaluate the effects on morphology, cell biology, gene expression, and relevant functions of different cell types that are resident in the gastrointestinal (GI) tract. An ideal near-physiological model should contain a mixture of different GI epithelial cells (e.g. Paneth cells, goblet cells, absorptive and hormone secretive epithelial cells), which can be cultured indefinitely. Recently, the culture and applications of long-term primary multi-cellular cluster structures gastrointestinal organoids (or enteroids) have been demonstrated, and within the last 5 years the number of researchers that commonly use this tissue culture model has increased rapidly. This multi-cellular system may be a promising addition for existing ex vivo and alternative for animal models for testing effects of food bioactives on the intestinal tissue, and could provide a model for pre-screening of compounds prior to moving to the large scale testing systems. Moreover, intestinal organoids can be cultured from different species (e.g. human, pig and mouse). In this chapter we will focus on organoids cultured from mouse and pig crypt cells. We will give a short overview on how to isolate, culture, incubate, and apply them in different research fields

Congenital diarrhoeal disorders: advances in this evolving web of inherited enteropathies

Congenital diarrhoeal disorders (CDDs) represent an evolving web of rare chronic enteropathies, with a typical onset early in life. In many of these conditions, severe chronic diarrhoea represents the primary clinical manifestation, whereas in others diarrhoea is only a component of a more complex multi-organ or systemic disorder. Typically, within the first days of life, diarrhoea leads to a life-threatening condition highlighted by severe dehydration and serum electrolyte abnormalities. Thus, in the vast majority of cases appropriate therapy must be started immediately to prevent dehydration and long-term, sometimes severe, complications. The number of well-characterized disorders attributed to CDDs has gradually increased over the past several years, and many new genes have been identified and functionally related to CDDs, opening new diagnostic and therapeutic perspectives. Molecular analysis has changed the diagnostic scenario in CDDs, and led to a reduction in invasive and expensive procedures. Major advances have been made in terms of pathogenesis, enabling a better understanding not only of these rare conditions but also of more common diseases mechanisms

Development and application of human adult stem or progenitor cell organoids

Adult stem or progenitor cell organoids are 3D adult-organ-derived epithelial structures that contain self-renewing and organ-specific stem or progenitor cells as well as differentiated cells. This organoid culture system was first established in murine intestine and subsequently developed for several other organs and translated to humans. Organoid cultures have proved valuable for basic research and for the study of healthy tissue homeostasis and the biology of disease. In addition, data from proof-of-principle experiments support promising clinical applications of adult stem or progenitor cell organoids. Although renal organoids have many potential applications, an adult stem or progenitor cell organoid culture system has not yet been developed for the kidney. The development of such a system is likely to be challenging because of the intricate renal architecture. Differentiated 3D cultures and stem or progenitor cell 3D sphere cultures are, however, available for the kidney. These cultures indicate the feasibility of renal organoid culture and provide a solid basis for its development. In this Review, we discuss the state-of-the-art of human adult stem or progenitor cell organoid culture and the potential of renal organoids as tools in basic and clinical research