Human iPS Cell-Derived Patient Tissues and 3D Cell Culture Part 2: Spheroids, Organoids, and Disease Modeling

Abstract

Human induced pluripotent stem cells (HiPSCs) provide several advantages for drug discovery, but principally they provide a source of clinically relevant tissue. Furthermore, the use of HiPSCs cultured in three-dimensional (3D) systems, as opposed to traditional two-dimensional (2D) culture approaches, better represents the complex tissue architecture in vivo. The use of HiPSCs in 3D spheroid and organoid culture is now growing, but particularly when using myocardial, intestinal enteric nervous system, and retinal cell lines. However, organoid cell culture is perhaps making the most notable impact in research and drug discovery, in which 3D neuronal cell cultures allow direct modeling of cortical cell layering and neuronal circuit activity. Given the specific degeneration seen in discrete neuronal circuitry in Alzheimer's disease (AD) and Parkinson's disease (PD), HiPSC culture systems are proving to be a major advance. In the present review, the second part of a two-part review, we discuss novel methods in which 3D cell culture systems (principally organoids) are now being used to provide insights into disease mechanisms. (The use of HiPSCs in target identification was reviewed in detail in Part 1.)

Modeling alcohol-induced neurotoxicity using human induced pluripotent stem cell-derived three-dimensional cerebral organoids

“…In 2013, Dr. Knoblich's lab developed an in vitro system of generating cerebral organoids by growing human iPSCs in Matrigel, a scaffold resembling the extracellular matrix, which allowed the cells to differentiate into cellular layers similar to those of real developing brains 36 . Recent data from us and others additionally support that cerebral organoids structurally and developmentally recapitulate fetal brains with higher fidelity than the currently used 2D brain cell models 36,40,[45][46][47][48][49][50][51] . Cerebral organoids have been used for modeling several human developmental brain diseases (e.g., microcephaly, autism, and Zika virus infection), and the findings have provided novel insights into these diseases' mechanisms and treatment 36,40,[52][53][54][55][56][57][58][59][60] .…”

Alzheimer’s Disease, and Breast and Prostate Cancer Research: Translational Failures and the Importance to Monitor Outputs and Impact of Funded Research

“…In particular, patient-derived cells, such as iPSCs, are often used to study human diseases in vitro (disease-in-a-dish models), providing insights into disease mechanisms. Such human cell-based models can be used along with human biological samples (e.g., blood, serum, tissue biopsies) for the definition of early biomarkers of human diseases and the design of new therapeutics [ 26 ]. AD patient-derived iPSC models harbor translational potential [ 27 ], and are already permitting the identification of AD biomarker candidates [ 28 ].…”

“…In an effort to develop and optimize more human relevant models and increase predictive capacity, a wide range of non-animal approaches have been developed in recent years, spanning from patient-derived cells, such as induced pluripotent stem cells (iPSCs) [26][27][28], three-dimensional (3-D) tumor spheroids [29], complex microfluidics organ-on-chip technologies [25,30], to next-generation sequencing and omics technologies, integrated computer modelling, systems biology, and imaging techniques [31,32]. These models and techniques, along with data derived from clinical and observational studies, can already be used in an integrated manner to gather insights into disease molecular and cellular mechanisms, discover new biomarkers, and design novel therapeutic and preventive strategies.…”

Factors to consider when interrogating 3D culture models with plate readers or automated microscopes

“…Recent advances have been made in optimizing the culture methods for using stem cells from normal and diseased patients to generate an unlimited supply of organoids containing multiple differentiated cell types closely reflecting in vivo biology. Researchers now can produce advanced 3D organoid models representing many human organs that are suitable for studying a variety of diseases in vitro (Aboulkheyr Es et al 2018;Artegiani and Clevers 2018;Eglen and Reisine 2019). These techniques overcome many of the disadvantages of using primary cells derived from patient biopsies, most notably heterogeneity of samples and the limited availability of source material.…”