Organoids are not organs: Sources of variation and misinformation in organoid biology

Jensen, Kim B.; Little, Melissa H.

doi:10.1016/j.stemcr.2023.05.009

Cited by 27 publications

(16 citation statements)

References 116 publications

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“…is not yet achieved, necessitating further research to create functional organs [8]. The other method of chimera organogenesis using blastocyst complementation, in which human pluripotent stem cells are injected into embryos genetically engineered not to develop specific organs, has also garnered attention [9][10][11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

“…On the other hand, studies on organogenesis utilizing human induced pluripotent stem cells (iPSCs) have also been actively pursued in recent years[5–7]. Nonetheless, the complete establishment of requisite cell types and environments for organogenesis is not yet achieved, necessitating further research to create functional organs[8]. The other method of chimera organogenesis using blastocyst complementation, in which human pluripotent stem cells are injected into embryos genetically engineered not to develop specific organs, has also garnered attention[9–14].…”

Section: Introductionmentioning

confidence: 99%

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Generation of human-pig chimeric renal organoids using iPSC technology

Fujimori,

Yamanaka,

Shimada

et al. 2024

Preprint

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The potential of using porcine organs and human induced pluripotent stem cell (iPSC)-derived organoids as alternative organs for human transplantation has garnered growing attention. However, both approaches still face technological challenges. Interspecies chimeric organ production using human iPSCs is expected to be another promising approach that addresses the challenges associated with organ production. Our research group successfully generated human-mouse chimeric renal organoids by utilizing human iPSC-derived nephron progenitor cells (NPCs) and fetal mouse kidneys. However, the current technology has limited engraftment and development capabilities for human NPCs, and there have been no reports of generating interspecies chimeric renal organoids in larger animals, limited only to rodents. Therefore, in this study, we embarked on the production of human-pig chimeric renal organoids using the pig kidney, which is considered the most promising source of organs for interspecies transplantation to humans. To construct a human-pig chimeric renal organoid culture system, we first modified the existing human-mouse chimeric renal organoid culture system and developed a method that enables the survival and continued renal development of both species. This method was found to be applicable to porcine fetal kidney cells, and ultimately, we successfully produced human-pig chimeric renal organoids. Furthermore, this culture method can also be applied to the generation of human interspecies chimeric kidneys for future clinical applications. The findings of this study serve as a foundational technology that will greatly accelerate future research in humanized pig kidney production for clinical purposes, and are also expected to be used as an evaluation technique to ensure the quality of human NPCs for xenotransplantation.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Generation of human-pig chimeric renal organoids using iPSC technology

Fujimori,

Yamanaka,

Shimada

et al. 2024

Preprint

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“…It is clear that BOs will continue to provide insight into human brain health and disease and reduce the use of animals in research. Yet, it is also clear that benchmarking human BOs, where ethically possible, to human brain tissue will be critical because species-specific protein processing often complicates the interpretation of data and its translation to the clinic ( Webster et al, 1995 ; Bellinger et al, 2011 ; Jensen and Little, 2023 ; Maksour and Ooi, 2023 ; Wenzel et al, 2023a ). Furthermore, the quality control of commercially available antibodies often relies on recombinant proteins or immortalized cell lines as positive controls; this is particularly important given that the protein banding pattern in a simplified system does not properly reflect the endogenous banding pattern in complex tissues such as brain parenchymal tissues and BOs, both of which have more multicellular machinery that could influence protein signatures ( Haytural et al, 2019 ; Rosell et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

Brain organoids engineered to give rise to glia and neural networks after 90 days in culture exhibit human-specific proteoforms

Wenzel,

Mousseau

2024

Front. Cell. Neurosci.

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Human brain organoids are emerging as translationally relevant models for the study of human brain health and disease. However, it remains to be shown whether human-specific protein processing is conserved in human brain organoids. Herein, we demonstrate that cell fate and composition of unguided brain organoids are dictated by culture conditions during embryoid body formation, and that culture conditions at this stage can be optimized to result in the presence of glia-associated proteins and neural network activity as early as three-months in vitro. Under these optimized conditions, unguided brain organoids generated from induced pluripotent stem cells (iPSCs) derived from male–female siblings are similar in growth rate, size, and total protein content, and exhibit minimal batch-to-batch variability in cell composition and metabolism. A comparison of neuronal, microglial, and macroglial (astrocyte and oligodendrocyte) markers reveals that profiles in these brain organoids are more similar to autopsied human cortical and cerebellar profiles than to those in mouse cortical samples, providing the first demonstration that human-specific protein processing is largely conserved in unguided brain organoids. Thus, our organoid protocol provides four major cell types that appear to process proteins in a manner very similar to the human brain, and they do so in half the time required by other protocols. This unique copy of the human brain and basic characteristics lay the foundation for future studies aiming to investigate human brain-specific protein patterning (e.g., isoforms, splice variants) as well as modulate glial and neuronal processes in an in situ-like environment.

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“…While human brain organoids (BOs) were first described over a decade ago (Eiraku et al, 2008), there is still a limited understanding about which features of the human brain these cultures mimic, and whether they can replace the use of animals to provide insight into health and disease (Wenzel et al, 2023). Benchmarking human BOs, where ethically possible, to human brain tissue is critical because interspecies differences complicate the interpretation of data (Bellinger et al, 2011; Jensen and Little, 2023; Maksour and Ooi, 2023; Webster et al, 1995). Furthermore, the quality control of antibodies by suppliers often relies on recombinant proteins or immortalized cell lines as positive controls for antibodies.…”

Section: Introductionmentioning

confidence: 99%

Protein expression profiles in brain organoids are more similar to those in human brain parenchyma than in mouse brain parenchyma

Wenzel,

Mousseau

2023

Preprint

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Human brain organoids are emerging as relevant models for the study of human brain health and disease. However, it has not been shown whether human brain organoids exhibit a proteoform profile similar to the human brain. Herein, we demonstrate that unguided brain organoids exhibit minimal batch-to-batch variability in cell composition and metabolism when generated from induced pluripotent stem cells (iPSCs) derived from male-female siblings. We then show that profiles of select proteins in these brain organoids are more similar to autopsied human cortical and cerebellar profiles than to those in mouse cortical samples. Brain organoids derived from sibling iPSCs do not exhibit any sex differences in protein proportions. By benchmarking human brain organoid proteoforms against human parenchymal tissue, we establish the foundation for future studies that could investigate, for example, how well brain organoids can model any of the known sex-dependent differences in cellular function, including responses of drug-receptor interactions.

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Organoids are not organs: Sources of variation and misinformation in organoid biology

Cited by 27 publications

References 116 publications

Generation of human-pig chimeric renal organoids using iPSC technology

Generation of human-pig chimeric renal organoids using iPSC technology

Brain organoids engineered to give rise to glia and neural networks after 90 days in culture exhibit human-specific proteoforms

Protein expression profiles in brain organoids are more similar to those in human brain parenchyma than in mouse brain parenchyma

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