Longitudinal morphological and functional characterization of human heart organoids using optical coherence tomography

Ming, Yixuan; Hao, Senyue; Wang, Fei; Lewis-Israeli, Yonatan R.; Volmert, Brett D.; Xu, Zhiyao; Goestenkors, Anna; Aguirre, Aitor; Zhou, Chao

doi:10.1016/j.bios.2022.114136

Cited by 33 publications

(20 citation statements)

References 42 publications

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“…Additionally, protocols for organoid generation and quality control are not globally standardized, affecting reproducibility, and their production and maintenance are relatively expensive compared to traditional cell lines or animal models. Finally, studies of organoids become increasingly difficult owing to the growing cellular and structural complexity required, and in many instances necessitate new technology development (Kim et al., 2020; Ming et al., 2022).…”

Section: Commentarymentioning

confidence: 99%

Modeling the Effects of Maternal Diabetes on the Developing Human Heart Using Pluripotent Stem Cell–Derived Heart Organoids

et al. 2022

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Congenital heart defects (CHD) constitute the most common type of birth defect in humans. Maternal diabetes during the first trimester of pregnancy (pregestational diabetes, or PGD) is one of the most prominent factors contributing to CHD, and is present in a significant population of female patients with diabetes in reproductive age. PGD is challenging to manage clinically due to the extreme sensitivity of the developing embryo to glucose oscillations, and constitutes a critical health problem for the mother and the fetus. The prevalence of PGD-induced CHD is increasing due to the ongoing diabetes epidemic. While studies using animal models and cells in culture have demonstrated that PGD alters critical cellular and developmental processes, the mechanisms remain obscure, and it is unclear to what extent these models recapitulate PGD-induced CHD in humans. Clinical practice precludes direct studies in developing human embryos, further highlighting the need for physiologically relevant models. To bypass many of these technical and ethical limitations, we describe here a human pluripotent stem cell (hPSC)-based method to generate developmentally relevant self-organizing human heart organoids. By using glucose and insulin to mimic the diabetic environment that the embryo faces in PGD, this system allows modeling critical features of PGD in a human system with relevant physiology, structure, and cell types. The protocol starts with the generation of hPSC-derived embryoid bodies in a 96-well plate, followed by a small molecule-based three-step Wnt activation/inhibition/activation strategy. Organoids are then differentiated under healthy (normal insulin and glucose) and diabetic conditions (high insulin and glucose) over time, allowing for the study of the effects of pregestational diabetes on the developing human heart. We also provide an immunofluorescence protocol for comparing, characterizing, and analyzing the differences between the healthy and diabetic organoids, and comment on additional steps for preparing the organoids for analysis by other techniques after differentiation.

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Section: Commentarymentioning

confidence: 99%

Modeling the Effects of Maternal Diabetes on the Developing Human Heart Using Pluripotent Stem Cell–Derived Heart Organoids

et al. 2022

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“…We decided to employ optical coherence tomography (OCT) to live image organoids over time, and to measure the growth and monitor dynamics of chamber development under developmental induction conditions via a custom-made OCT microscopy system amenable to high-content screening 114, 115 (Fig. 8a) .…”

Section: Resultsmentioning

confidence: 99%

“…A Spectral-Domain Optical Coherence Tomography (SD-OCT) system similar to our previous work 114, 115 was used for label-free longitudinal imaging of the heart organoids. A superluminescent diode (EXALOS, EXC250023-00) was used as the light source with a center wavelength of ∼1300 nm and a 3 dB spectrum range of ∼180 nm.…”

Section: Methodsmentioning

confidence: 99%

A patterned human heart tube organoid model generated by pluripotent stem cell self-assembly

Volmert

Aguirre

Riggs

et al. 2022

Preprint

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Human pluripotent stem cells can recapitulate significant features of mammalian organ development in vitro, including key aspects of heart development. We hypothesized that the organoids thus created can be made substantially more relevant by mimicking aspects of in utero gestation, leading to higher physiological and anatomical resemblance to their in vivo counterparts. Here, we report steps towards generating developmentally inspired maturation methodologies to differentiate early human heart organoids into patterned heart-tube-like structures in a reproducible and high-throughput fashion by complete self-organization. The maturation strategy consists of the controlled and stepwise exposure to metabolic (glucose, fatty acids) and hormonal signals (T3, IGF-1) as present during early heart development. These conditions elicit important transcriptomic, cellular, morphological, metabolomic, and functional changes over a 10-day period consistent with continuously increasing heart complexity, maturation, and patterning. Our data reveals the emergence of atrial and ventricular cardiomyocyte populations, valvular cells, epicardial cells, proepicardial-derived cells, endothelial cells, stromal cells, conductance cells, and cardiac progenitors, all of them cell types present in the primitive heart tube. Anatomically, the organoids elongate and develop well-differentiated atrial and ventricular chambers with compacted myocardial muscle walls and a proepicardial organ. For the first time in a completely self-organizing heart organoid, we show anterior-posterior patterning due to an endogenous retinoic acid gradient originating at the atrial pole, where proepicardial and atrial populations reside, mimicking the developmental process present within the primitive heart tube. Collectively, these findings highlight the ability of self-organization and developmental maturation strategies to recapitulate human heart development. Our patterned human heart tube model constitutes a powerful in vitro tool for dissecting the role of different cell types and genes in human heart development, as well as disease modeling congenital heart defects, and represents a step forward in creating fully synthetic human hearts.

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“…Various factors such as variation in overall morphology and the cell ratio can indicate whether the organoids successfully resemble their in vivo counterparts or the morphological changes to external stimuli. [ 7 ] However, improved genetic technologies enabled the engineered organoids to closely mimic both the morphology and the functional ability of their counterparts. Since organoids show dynamic responses, the need to monitor their activities was apparent.…”

Section: Introductionmentioning

confidence: 99%

Recent Advances in Electrophysiological Recording Platforms for Brain and Heart Organoids

Chung

Kim

Song

et al. 2022

Advanced NanoBiomed Research

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Organoids refer to 3D stem cells that have been developed to model neurological disorders in vitro. Typically, brain and heart organoids have gained interest for their potential to truly mimic the functional ability of real organs. Morphological analysis methods using immunostaining and slicing of the organoids are explored extensively over the past decade to evaluate the structures and functions of organoids. However, the destructiveness of these methods limits real‐time monitoring of the dynamic responses of the organoids. Therefore, electrophysiological functional analysis of organoids with minimally invasive forms can be a key solution to an improved understanding of the nature of complex organoids. Herein, the latest advances in the recording platforms are reviewed and a comprehensive study of considerations regarding electrophysiological factors is provided. Furthermore, current challenges are discussed along with prospects for next‐generation electrophysiological analysis of brain and heart organoids.

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Longitudinal morphological and functional characterization of human heart organoids using optical coherence tomography

Cited by 33 publications

References 42 publications

Modeling the Effects of Maternal Diabetes on the Developing Human Heart Using Pluripotent Stem Cell–Derived Heart Organoids

Modeling the Effects of Maternal Diabetes on the Developing Human Heart Using Pluripotent Stem Cell–Derived Heart Organoids

A patterned human heart tube organoid model generated by pluripotent stem cell self-assembly

Recent Advances in Electrophysiological Recording Platforms for Brain and Heart Organoids

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