Gastruloids enable modeling of the earliest stages of human cardiac and hepatic vascularization.
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| Title: | Gastruloids enable modeling of the earliest stages of human cardiac and hepatic vascularization. |
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| Authors: | Abilez, Oscar J., Yang, Huaxiao, Guan, Yuan, Shen, Mengcheng, Yildirim, Zehra, Zhuge, Yan, Venkateshappa, Ravichandra, Zhao, Shane R., Gomez, Angello H., El-Mokahal, Marcel, Dunkenberger, Logan, Ono, Yoshikazu, Shibata, Masafumi, Nwokoye, Peter N., Tian, Lei, Wilson, Kitchener D., Lyall, Evan H., Jia, Fangjun, Wo, Hung Ta, Zhou, Gao |
| Source: | Science. 6/5/2025, Vol. 388 Issue 6751, p1-23. 23p. |
| Subjects: | Heart, Embryos, Pluripotent stem cells, Organoids, Developmental biology |
| Abstract: | Although model organisms have provided insight into the earliest stages of cardiac and hepatic vascularization, we know very little about this process in humans because of ethical restrictions and the technical difficulty of obtaining embryos during very early development. In this study, we demonstrate that micropatterned human pluripotent stem cell–derived gastruloids enable in vitro modeling of the earliest stages of vascularization. We identify a combination of vascular-inducing factors that give rise to cardiac vascularized organoids with a spatially organized and branched vascular network. To show the broader utility of our vascularization strategy, we use the same vascular-inducing factors to produce hepatic vascularized organoids. Our results suggest that a conserved developmental program generates the vasculature within different types of organs. Editor's summary: Monitoring the steps of mammalian development is challenging at best and often impossible to do in human embryos. Organoids made from human-derived cells offer a tractable alternative, but they often lack key cell types that are present in an intact organism and may not properly mimic the cellular microenvironment seen during physiological development. By testing numerous differentiation conditions, Abilez et al. developed a method of culturing human pluripotent stem cells that differentiated into vascularized cardiac and hepatic organoids. These vascularized organoids could be used for studying cardiac and hepatic development and for addressing more immediately practical questions, such as the impact of drug exposure on human organ development. —Yevgeniya Nusinovich INTRODUCTION: Human pluripotent stem cells (hPSCs), including human embryonic stem cells and human induced pluripotent stem cells, can differentiate into various cell types of the body, such as cardiomyocytes, hepatocytes, and individual types of vascular cells. Furthermore, hPSCs can be used to create organoids, which are self-organizing three-dimensional (3D) structures that mimic key structural and functional characteristics of their in vivo organ counterparts. Several approaches to create organoids with a vascular system have been pursued to (i) avoid necrosis in the center of organoids where oxygen tension is low; (ii) achieve larger organoid growth for improving fidelity in modeling development, modeling diseases, and discovering new drugs; and (iii) increase the viability of implanted organoids used as regenerative therapies. However, codifferentiating the key cell types of an organoid along with a de novo vasculature comprising robust branching, hierarchical organization, and lumina formation has not been fully achieved. RATIONALE: Although model organisms have provided insight into the earliest stages of organ vascularization, we know very little about this process in humans because of ethical restrictions and the technical difficulty of obtaining human embryos at early developmental stages. However, hPSCs have been shown to model key aspects of development, including primitive streak formation, gastrulation, germ layer formation, and individual organ-specific cell type creation. Moreover, geometric micropatterning of hPSCs has enabled reproducible and scalable modeling of these developmental processes. With this basis, we developed an in vitro model to mimic the earliest developmental stages of cardiac and hepatic organoid vascularization, which corresponds to the first 3 weeks of in vivo human development and Carnegie Stages 9 and 10. RESULTS: Using four hPSC fluorescent reporter systems and spatially micropatterned hPSCs, we produced cardiac vascularized organoids (cVOs) in a scalable and reproducible fashion. Notably, we used the four reporter systems to characterize gastruloid, progenitor, and cardiovascular cell type formation in situ in developing cVOs. We identified a growth factor and small molecule cocktail that when added to micropatterned hPSCs generated a spatially organized, branched, and lumenized vascular network within a multilineage cVO comprising endocardial, myocardial, epicardial, and neuronal cell types. Single-cell transcriptomics, high-resolution 3D microscopy, and multiple functional analyses showed that cVOs were structurally and functionally similar to that of a 6.5–postconception week human embryonic heart at Carnegie Stages 19 and 20; however, our comparison also revealed differences that warrant future investigation. Additionally, we found that NOTCH and bone morphogenetic protein (BMP) signaling were required for vascularization in cVOs, with BMP inhibition having a more negative effect on vascular formation than NOTCH. To demonstrate the broader utility of our vascularization strategy, we used the same vascular-inducing cocktail to produce hepatic vascularized organoids (hVOs), which also comprised a spatially organized, branched, and lumenized vascular network integrated with multilineage hepatic cell types. CONCLUSION: Our in vitro model represents a technical advance for addressing questions regarding de novo organ vascularization. Furthermore, our results suggest that a conserved developmental program is involved in creating the vasculature within different organ systems. Micropatterning of four hPSC reporter lines enabled gastruloid, cardiovascular, progenitor, and cVO formation.: The identification of a vascular-inducing cocktail of growth factors enabled generation of a spatially organized, branched, and lumenized vascular network within a multilineage cVO. Transcriptomics, high-resolution 3D microscopy, and functional analyses showed that cVOs are similar to a human embryonic heart. The same vascular-inducing cocktail was then used to produce hVOs. CMs, cardiomyocytes; ECs, endothelial cells; SMCs, smooth muscle cells; PCs, pericytes; HCs, hepatocytes. [ABSTRACT FROM AUTHOR] |
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| Database: | Psychology and Behavioral Sciences Collection |
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| Abstract: | Although model organisms have provided insight into the earliest stages of cardiac and hepatic vascularization, we know very little about this process in humans because of ethical restrictions and the technical difficulty of obtaining embryos during very early development. In this study, we demonstrate that micropatterned human pluripotent stem cell–derived gastruloids enable in vitro modeling of the earliest stages of vascularization. We identify a combination of vascular-inducing factors that give rise to cardiac vascularized organoids with a spatially organized and branched vascular network. To show the broader utility of our vascularization strategy, we use the same vascular-inducing factors to produce hepatic vascularized organoids. Our results suggest that a conserved developmental program generates the vasculature within different types of organs. Editor's summary: Monitoring the steps of mammalian development is challenging at best and often impossible to do in human embryos. Organoids made from human-derived cells offer a tractable alternative, but they often lack key cell types that are present in an intact organism and may not properly mimic the cellular microenvironment seen during physiological development. By testing numerous differentiation conditions, Abilez et al. developed a method of culturing human pluripotent stem cells that differentiated into vascularized cardiac and hepatic organoids. These vascularized organoids could be used for studying cardiac and hepatic development and for addressing more immediately practical questions, such as the impact of drug exposure on human organ development. —Yevgeniya Nusinovich INTRODUCTION: Human pluripotent stem cells (hPSCs), including human embryonic stem cells and human induced pluripotent stem cells, can differentiate into various cell types of the body, such as cardiomyocytes, hepatocytes, and individual types of vascular cells. Furthermore, hPSCs can be used to create organoids, which are self-organizing three-dimensional (3D) structures that mimic key structural and functional characteristics of their in vivo organ counterparts. Several approaches to create organoids with a vascular system have been pursued to (i) avoid necrosis in the center of organoids where oxygen tension is low; (ii) achieve larger organoid growth for improving fidelity in modeling development, modeling diseases, and discovering new drugs; and (iii) increase the viability of implanted organoids used as regenerative therapies. However, codifferentiating the key cell types of an organoid along with a de novo vasculature comprising robust branching, hierarchical organization, and lumina formation has not been fully achieved. RATIONALE: Although model organisms have provided insight into the earliest stages of organ vascularization, we know very little about this process in humans because of ethical restrictions and the technical difficulty of obtaining human embryos at early developmental stages. However, hPSCs have been shown to model key aspects of development, including primitive streak formation, gastrulation, germ layer formation, and individual organ-specific cell type creation. Moreover, geometric micropatterning of hPSCs has enabled reproducible and scalable modeling of these developmental processes. With this basis, we developed an in vitro model to mimic the earliest developmental stages of cardiac and hepatic organoid vascularization, which corresponds to the first 3 weeks of in vivo human development and Carnegie Stages 9 and 10. RESULTS: Using four hPSC fluorescent reporter systems and spatially micropatterned hPSCs, we produced cardiac vascularized organoids (cVOs) in a scalable and reproducible fashion. Notably, we used the four reporter systems to characterize gastruloid, progenitor, and cardiovascular cell type formation in situ in developing cVOs. We identified a growth factor and small molecule cocktail that when added to micropatterned hPSCs generated a spatially organized, branched, and lumenized vascular network within a multilineage cVO comprising endocardial, myocardial, epicardial, and neuronal cell types. Single-cell transcriptomics, high-resolution 3D microscopy, and multiple functional analyses showed that cVOs were structurally and functionally similar to that of a 6.5–postconception week human embryonic heart at Carnegie Stages 19 and 20; however, our comparison also revealed differences that warrant future investigation. Additionally, we found that NOTCH and bone morphogenetic protein (BMP) signaling were required for vascularization in cVOs, with BMP inhibition having a more negative effect on vascular formation than NOTCH. To demonstrate the broader utility of our vascularization strategy, we used the same vascular-inducing cocktail to produce hepatic vascularized organoids (hVOs), which also comprised a spatially organized, branched, and lumenized vascular network integrated with multilineage hepatic cell types. CONCLUSION: Our in vitro model represents a technical advance for addressing questions regarding de novo organ vascularization. Furthermore, our results suggest that a conserved developmental program is involved in creating the vasculature within different organ systems. Micropatterning of four hPSC reporter lines enabled gastruloid, cardiovascular, progenitor, and cVO formation.: The identification of a vascular-inducing cocktail of growth factors enabled generation of a spatially organized, branched, and lumenized vascular network within a multilineage cVO. Transcriptomics, high-resolution 3D microscopy, and functional analyses showed that cVOs are similar to a human embryonic heart. The same vascular-inducing cocktail was then used to produce hVOs. CMs, cardiomyocytes; ECs, endothelial cells; SMCs, smooth muscle cells; PCs, pericytes; HCs, hepatocytes. [ABSTRACT FROM AUTHOR] |
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| ISSN: | 00368075 |
| DOI: | 10.1126/science.adu9375 |
