![]() ![]() Excess blood can be removed by immersing tissue in sterile phosphate-buffered saline prior to transfer into a sterile petri-dish for dissection into <1 mm 3 fragments using two sterilized carbon steel single-edged razor blades (T585 Agar Scientific, UK) stuck together under a dissecting microscope (to aid in sample selection and orientation). ![]() “Time zero” samples are processed concomitant with the onset of organ culture to establish morphological features prior to culture and treatments. Tissue for HOC (Figure 1) is transported to the laboratory as quickly as possible to minimize deterioration, ideally within minutes of collection. Methodology of HOC The Method Described Here Reflects Our Approach for Optimizing HOC ![]() More recent examples have illustrated how newer morphological approaches, such as in situ proximity ligation assay, can be applied to HOCs to reveal greater details in specific biological responses. To do so, we have chosen illustrative examples from the literature from the past 30 years that have been used to gain insights into inflammatory process and diseases, cancer, and stem cell biology. In this review, we will illustrate how human organ cultures (HOCs) offer a simple approach that may better address these issues. Newer culture approaches, such as three-dimensional (3D) cultures, organoids, or organs-on-a-chip have attempted to better replicate the tissue microenvironment, but have been only partly successful. Traditional human cell cultures, which are often used to account for species differences are also limited in their representation of in vivo responses due to lack of an appropriate microenvironmental context of the responding cell types. Preclinical animal studies have had only limited success in predicting human physiology, pathology, and therapeutic responses. ![]() Examples are provided involving use of HOCs to study inflammation, cancer, and stem cell biology. We will specifically describe how HOCs can be combined with both traditional and more modern morphological techniques to reveal how anatomic location can alter cellular responses at a molecular level and permit comparisons among different cells and different cell types within the same tissue. Here, we review how human organ cultures (HOCs) can more faithfully preserve in vivo tissue architecture and can better represent disease-associated changes. Three-dimensional cultures that position cells on synthetic matrices, or organoid or organ-on-a-chip cultures, in which different cell spontaneously organize contacts with other cells and natural matrix only partly overcome this limitation. Standard human cell cultures can address some of these concerns but the absence of the normal tissue microenvironment can alter cellular responses. Human studies, critical for developing new diagnostics and therapeutics, are limited by ethical and logistical issues, and preclinical animal studies are often poor predictors of human responses. 2Department of Immunobiology, Yale University School of Medicine, New Haven, CT, United States.1Department of Medicine, NIHR Cambridge Biomedical Research Centre, University of Cambridge, Cambridge, United Kingdom. ![]()
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