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1.
Life Sci ; 271: 119195, 2021 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-33581125

RESUMO

AIMS: Ulcerative colitis and Crohn's disease, collectively known as inflammatory bowel disease (IBD), are chronic inflammatory disorders of the intestine for which key elements in disease initiation and perpetuation are defects in epithelial barrier integrity. Achieving mucosal healing is essential to ameliorate disease outcome and so new therapies leading to epithelial homeostasis and repair are under investigation. This study was designed to determine the mechanisms by which IL-22 regulates intestinal epithelial cell function. MAIN METHODS: Human intestinal organoids and resections, as well as mice were used to evaluate the effect of IL-22 on stem cell expansion, proliferation and expression of mucus components. IL-22 effect on barrier function was assessed in polarized T-84 cell monolayers. Butyrate co-treatments and organoid co-cultures with immune cells were performed to monitor the impact of microbial-derived metabolites and inflammatory environments on IL-22 responses. KEY FINDINGS: IL-22 led to epithelial stem cell expansion, proliferation, barrier dysfunction and anti-microbial peptide production in human and mouse models evaluated. IL-22 also altered the mucus layer by inducing an increase in membrane mucus but a decrease in secreted mucus and goblet cell content. IL-22 had the same effect on anti-microbial peptides and membrane mucus in both healthy and IBD human samples. In contrast, this IL-22-associated epithelial phenotype was different when treatments were performed in presence of butyrate and organoids co-cultured with immune cells. SIGNIFICANCE: Our data indicate that IL-22 promotes epithelial regeneration, innate defense and membrane mucus production, strongly supporting the potential clinical utility of IL-22 as a mucosal healing therapy in IBD.


Assuntos
Células Epiteliais/fisiologia , Homeostase/fisiologia , Interleucinas/fisiologia , Interleucinas/uso terapêutico , Mucosa Intestinal/fisiologia , Animais , Linhagem Celular , Técnicas de Cocultura , Colite Ulcerativa/tratamento farmacológico , Colite Ulcerativa/patologia , Células Epiteliais/efeitos dos fármacos , Homeostase/efeitos dos fármacos , Humanos , Interleucinas/farmacologia , Mucosa Intestinal/citologia , Mucosa Intestinal/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Organoides/efeitos dos fármacos , Organoides/fisiologia
2.
Science ; 371(6531): 839-846, 2021 02 19.
Artigo em Inglês | MEDLINE | ID: mdl-33602855

RESUMO

Organoid technology holds great promise for regenerative medicine but has not yet been applied to humans. We address this challenge using cholangiocyte organoids in the context of cholangiopathies, which represent a key reason for liver transplantation. Using single-cell RNA sequencing, we show that primary human cholangiocytes display transcriptional diversity that is lost in organoid culture. However, cholangiocyte organoids remain plastic and resume their in vivo signatures when transplanted back in the biliary tree. We then utilize a model of cell engraftment in human livers undergoing ex vivo normothermic perfusion to demonstrate that this property allows extrahepatic organoids to repair human intrahepatic ducts after transplantation. Our results provide proof of principle that cholangiocyte organoids can be used to repair human biliary epithelium.


Assuntos
Doenças dos Ductos Biliares/terapia , Ductos Biliares Intra-Hepáticos/fisiologia , Ductos Biliares/citologia , Terapia Baseada em Transplante de Células e Tecidos , Células Epiteliais/citologia , Organoides/transplante , Animais , Bile , Ductos Biliares/fisiologia , Ductos Biliares Intra-Hepáticos/citologia , Ducto Colédoco/citologia , Células Epiteliais/fisiologia , Vesícula Biliar/citologia , Regulação da Expressão Gênica , Humanos , Fígado/fisiologia , Transplante de Fígado , Transplante de Células-Tronco Mesenquimais , Camundongos , Organoides/fisiologia , RNA-Seq , Obtenção de Tecidos e Órgãos , Transcriptoma
4.
Nat Biomed Eng ; 5(1): 11-25, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33318650

RESUMO

Engineered human mini-brains, made possible by knowledge from the convergence of precision microengineering and cell biology, permit systematic studies of complex neurological processes and of pathogenesis beyond what can be done with animal models. By culturing human brain cells with physiological microenvironmental cues, human mini-brain models reconstitute the arrangement of structural tissues and some of the complex biological functions of the human brain. In this Review, we highlight the most significant developments that have led to microphysiological human mini-brain models. We introduce the history of mini-brain development, review methods for creating mini-brain models in static conditions, and discuss relevant state-of-the-art dynamic cell-culture systems. We also review human mini-brain models that reconstruct aspects of major neurological disorders under static or dynamic conditions. Engineered human mini-brains will contribute to advancing the study of the physiology and aetiology of neurological disorders, and to the development of personalized medicines for them.


Assuntos
Encéfalo , Modelos Biológicos , Organoides , Técnicas de Cultura de Tecidos , Doença de Alzheimer , Encéfalo/anatomia & histologia , Encéfalo/citologia , Encéfalo/efeitos dos fármacos , Encéfalo/fisiologia , Lesões Encefálicas Traumáticas , Neoplasias Encefálicas , Técnicas de Cultura de Células , Células Cultivadas , Humanos , Miniaturização , Organoides/anatomia & histologia , Organoides/citologia , Organoides/efeitos dos fármacos , Organoides/fisiologia
5.
Neuron ; 107(6): 1014-1028, 2020 09 23.
Artigo em Inglês | MEDLINE | ID: mdl-32970996

RESUMO

The recent advent of human pluripotent stem cell (PSC)-derived 3D brain organoids has opened a window into aspects of human brain development that were not accessible before, allowing tractable monitoring and assessment of early developmental processes. However, their broad and effective use for modeling later stages of human brain development and disease is hampered by the lack of a stereotypic anatomical organization, which limits maturation processes dependent upon formation of unique cellular interactions and short- and long-range network connectivity. Emerging methods and technologies aimed at tighter regulatory control through bioengineering approaches, along with newer unbiased organoid analysis readouts, should resolve several of the current limitations. Here, we review recent advances in brain organoid generation and characterization with a focus on highlighting future directions utilizing interdisciplinary strategies that will be important for improving the physiological relevance of this model system.


Assuntos
Encéfalo/citologia , Crescimento Neuronal , Organoides/citologia , Cultura Primária de Células/métodos , Encéfalo/metabolismo , Encéfalo/fisiologia , Genômica/métodos , Humanos , Células-Tronco Neurais/citologia , Células-Tronco Neurais/metabolismo , Células-Tronco Neurais/fisiologia , Organoides/metabolismo , Organoides/fisiologia
6.
Nat Commun ; 11(1): 3791, 2020 07 29.
Artigo em Inglês | MEDLINE | ID: mdl-32728089

RESUMO

Brain organoids are promising tools for disease modeling and drug development. For proper neuronal network formation excitatory and inhibitory neurons as well as glia need to co-develop. Here, we report the directed self-organization of human induced pluripotent stem cells in a collagen hydrogel towards a highly interconnected neuronal network at a macroscale tissue format. Bioengineered Neuronal Organoids (BENOs) comprise interconnected excitatory and inhibitory neurons with supportive astrocytes and oligodendrocytes. Giant depolarizing potential (GDP)-like events observed in early BENO cultures mimic early network activity of the fetal brain. The observed GABA polarity switch and reduced GDPs in >40 day BENO indicate progressive neuronal network maturation. BENOs demonstrate expedited complex network burst development after two months and evidence for long-term potentiation. The similarity of structural and functional properties to the fetal brain may allow for the application of BENOs in studies of neuronal plasticity and modeling of disease.


Assuntos
Encéfalo/citologia , Neurogênese , Plasticidade Neuronal/fisiologia , Organoides/fisiologia , Engenharia Tecidual/métodos , Potenciais de Ação/fisiologia , Encéfalo/crescimento & desenvolvimento , Técnicas de Cultura de Células , Diferenciação Celular , Humanos , Células-Tronco Pluripotentes Induzidas/fisiologia , Neurônios/fisiologia , Ácido gama-Aminobutírico/metabolismo
7.
Nat Rev Mol Cell Biol ; 21(10): 571-584, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32636524

RESUMO

The historical reliance of biological research on the use of animal models has sometimes made it challenging to address questions that are specific to the understanding of human biology and disease. But with the advent of human organoids - which are stem cell-derived 3D culture systems - it is now possible to re-create the architecture and physiology of human organs in remarkable detail. Human organoids provide unique opportunities for the study of human disease and complement animal models. Human organoids have been used to study infectious diseases, genetic disorders and cancers through the genetic engineering of human stem cells, as well as directly when organoids are generated from patient biopsy samples. This Review discusses the applications, advantages and disadvantages of human organoids as models of development and disease and outlines the challenges that have to be overcome for organoids to be able to substantially reduce the need for animal experiments.


Assuntos
Biologia/métodos , Medicina/métodos , Organoides/fisiologia , Animais , Doenças Transmissíveis/patologia , Doenças Genéticas Inatas/patologia , Engenharia Genética/métodos , Humanos , Neoplasias/patologia , Células-Tronco/fisiologia
8.
Med Sci (Paris) ; 36(6-7): 600-606, 2020.
Artigo em Francês | MEDLINE | ID: mdl-32614311

RESUMO

In inherited retinal diseases such retinitis pigmentosa, characterized by progressive loss of light sensitive neurons (photoreceptors), cell therapy is now considered as an attractive strategy. Photoreceptor cell replacement would be valuable for restoring function to retinas in a way that is independent from the cause of the disease. With advances in stem cell biology, considerable strides have been made towards the generation of retinal cells, in particular with the development of 3D culture systems allowing the generation of retinal organoids from pluripotent stem cells. In this review, we present a state-of-the art of preclinical strategies conducted in animal models for photoreceptor replacement from stem cell-derived photoreceptors and we discuss the important obstacles to overcome in the future.


Assuntos
Células Fotorreceptoras/transplante , Retinite Pigmentosa/terapia , Terapias em Estudo/métodos , Terapias em Estudo/tendências , Animais , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/transplante , Organoides/citologia , Organoides/fisiologia , Células Fotorreceptoras/citologia , Células Fotorreceptoras/fisiologia , Células-Tronco Pluripotentes/citologia , Células-Tronco Pluripotentes/transplante , Retina/citologia , Retina/embriologia , Retina/transplante , Degeneração Retiniana/terapia , Retinite Pigmentosa/patologia , Índice de Gravidade de Doença , Técnicas de Cultura de Tecidos , Coleta de Tecidos e Órgãos/métodos , Coleta de Tecidos e Órgãos/normas , Coleta de Tecidos e Órgãos/tendências
9.
Med Sci (Paris) ; 36(6-7): 626-632, 2020.
Artigo em Francês | MEDLINE | ID: mdl-32614314

RESUMO

Generation of retinal organoids from pluripotent stem cells represents an important advance in the study of retinal development and offer new perspectives for the study of retinal diseases missing suitable animal models. Understanding the key stages of retinal development in vertebrates enabled to design protocols to generate self-organized three-dimensional structures derived from pluripotent stem cells and containing all retinal cell types. In addition to their application in basic research, such as the characterization of molecular and cellular mechanisms in retinal pathophysiology, these miniature organs also open up encouraging prospects in the field of cell therapy or the screening of therapeutic molecules, although some obstacles remain to be overcome.


Assuntos
Organoides/citologia , Retina/citologia , Doenças Retinianas/etiologia , Doenças Retinianas/patologia , Doenças Retinianas/terapia , Animais , Células Cultivadas , Humanos , Modelos Biológicos , Organoides/fisiologia , Retina/patologia , Retina/fisiologia , Terapias em Estudo/métodos , Terapias em Estudo/tendências , Técnicas de Cultura de Tecidos/métodos , Técnicas de Cultura de Tecidos/tendências
10.
Science ; 369(6500)2020 07 10.
Artigo em Inglês | MEDLINE | ID: mdl-32527923

RESUMO

Cerebrospinal fluid (CSF) is a vital liquid, providing nutrients and signaling molecules and clearing out toxic by-products from the brain. The CSF is produced by the choroid plexus (ChP), a protective epithelial barrier that also prevents free entry of toxic molecules or drugs from the blood. Here, we establish human ChP organoids with a selective barrier and CSF-like fluid secretion in self-contained compartments. We show that this in vitro barrier exhibits the same selectivity to small molecules as the ChP in vivo and that ChP-CSF organoids can predict central nervous system (CNS) permeability of new compounds. The transcriptomic and proteomic signatures of ChP-CSF organoids reveal a high degree of similarity to the ChP in vivo. Finally, the intersection of single-cell transcriptomics and proteomic analysis uncovers key human CSF components produced by previously unidentified specialized epithelial subtypes.


Assuntos
Barreira Hematoencefálica/fisiologia , Líquido Cefalorraquidiano/fisiologia , Plexo Corióideo/fisiologia , Organoides/fisiologia , Técnicas de Cultura de Células , Líquido Cefalorraquidiano/metabolismo , Proteínas do Líquido Cefalorraquidiano/metabolismo , Perfilação da Expressão Gênica , Humanos , Proteômica , Análise de Célula Única
11.
PLoS One ; 15(6): e0233860, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32479513

RESUMO

The generation of laminated and light responsive retinal organoids from induced pluripotent stem cells (iPSCs) provides a powerful tool for the study of retinal diseases and drug discovery and a robust platform for cell-based therapies. The aim of this study is to investigate whether retinal organoids can retain their morphological and functional characteristics upon storage at room temperature (RT) conditions and shipment by air using a commercially available container that maintains the environment at ambient temperature. Morphological analysis and measurements of neuroepithelial thickness revealed no differences between control, RT incubated and shipped organoids. Similarly immunohistochemical analysis showed no differences in cell type composition and position within the laminated retinal structure. All groups showed a similar response to light, suggesting that the biological function of retinal organoids was not affected by RT storage or shipment. These findings provide an advance in transport of ready-made retinal organoids, increasing their availability to many research and pharma labs worldwide and facilitating cross-collaborative research.


Assuntos
Organoides/transplante , Serviços Postais , Retina/citologia , Doenças Retinianas/terapia , Diferenciação Celular , Linhagem Celular , Avaliação Pré-Clínica de Medicamentos/métodos , Humanos , Células-Tronco Pluripotentes Induzidas/fisiologia , Luz , Organoides/efeitos dos fármacos , Organoides/fisiologia , Organoides/efeitos da radiação , Temperatura
12.
Science ; 368(6490): 497-505, 2020 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-32355025

RESUMO

Androgen deprivation is the cornerstone of prostate cancer treatment. It results in involution of the normal gland to ~90% of its original size because of the loss of luminal cells. The prostate regenerates when androgen is restored, a process postulated to involve stem cells. Using single-cell RNA sequencing, we identified a rare luminal population in the mouse prostate that expresses stemlike genes (Sca1 + and Psca +) and a large population of differentiated cells (Nkx3.1 +, Pbsn +). In organoids and in mice, both populations contribute equally to prostate regeneration, partly through androgen-driven expression of growth factors (Nrg2, Rspo3) by mesenchymal cells acting in a paracrine fashion on luminal cells. Analysis of human prostate tissue revealed similar differentiated and stemlike luminal subpopulations that likewise acquire enhanced regenerative potential after androgen ablation. We propose that prostate regeneration is driven by nearly all persisting luminal cells, not just by rare stem cells.


Assuntos
Androgênios/metabolismo , Próstata/fisiologia , Próstata/cirurgia , Neoplasias da Próstata/cirurgia , Regeneração , Antagonistas de Androgênios/uso terapêutico , Proteína de Ligação a Androgênios/genética , Animais , Antígenos de Neoplasias/genética , Ataxina-1/genética , Diferenciação Celular/genética , Proteínas Ligadas por GPI/genética , Expressão Gênica , Proteínas de Homeodomínio/genética , Humanos , Masculino , Células-Tronco Mesenquimais/fisiologia , Camundongos , Proteínas de Neoplasias/genética , Fatores de Crescimento Neural/genética , Tamanho do Órgão , Organoides/metabolismo , Organoides/fisiologia , Próstata/metabolismo , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/metabolismo , Regeneração/genética , Análise de Sequência de RNA , Análise de Célula Única , Trombospondinas/genética , Fatores de Transcrição/genética
13.
Med Sci (Paris) ; 36(4): 382-388, 2020 Apr.
Artigo em Francês | MEDLINE | ID: mdl-32356715

RESUMO

As burden of chronic respiratory diseases is constantly increasing, improving in vitro lung models is essential in order to reproduce as closely as possible the complex pulmonary architecture, responsible for oxygen uptake and carbon dioxide clearance. The study of diseases that affect the respiratory system has benefited from in vitro reconstructions of the respiratory epithelium with inserts in air/liquid interface (2D) or in organoids able to mimic up to the arborescence of the respiratory tree (3D). Recent development in the fields of pluripotent stem cells-derived organoids and genome editing technologies has provided new insights to better understand pulmonary diseases and to find new therapeutic perspectives.


Assuntos
Técnicas de Cultura de Células , Pulmão/citologia , Organoides/citologia , Células Epiteliais Alveolares/citologia , Células Epiteliais Alveolares/fisiologia , Animais , Bioengenharia/métodos , Bioengenharia/tendências , Dióxido de Carbono/farmacologia , Dióxido de Carbono/fisiologia , Técnicas de Cultura de Células/métodos , Técnicas de Cultura de Células/tendências , Células Cultivadas , Edição de Genes/métodos , Edição de Genes/tendências , Humanos , Pulmão/patologia , Pulmão/fisiologia , Modelos Biológicos , Organoides/patologia , Organoides/fisiologia , Oxigênio/farmacologia , Oxigênio/fisiologia , Troca Gasosa Pulmonar/fisiologia , Mucosa Respiratória/citologia , Mucosa Respiratória/efeitos dos fármacos , Tecidos Suporte/química
14.
Am J Physiol Cell Physiol ; 319(1): C151-C165, 2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32459504

RESUMO

In vitro cell cultures are crucial research tools for modeling human development and diseases. Although the conventional monolayer cell cultures have been widely used in the past, the lack of tissue architecture and complexity of such model fails to inform the true biological processes in vivo. Recent advances in the organoid technology have revolutionized the in vitro culture tools for biomedical research by creating powerful three-dimensional (3D) models to recapitulate the cellular heterogeneity, structure, and functions of the primary tissues. Such organoid technology enables researchers to recreate human organs and diseases in a dish and thus holds great promises for many translational applications such as regenerative medicine, drug discovery, and precision medicine. In this review, we provide an overview of the organoid history and development. We discuss the strengths and limitations of organoids as well as their potential applications in the laboratory and the clinic.


Assuntos
Pesquisa Biomédica/métodos , Técnicas de Cultura de Células/métodos , Modelos Biológicos , Organoides/fisiologia , Animais , Pesquisa Biomédica/tendências , Técnicas de Cultura de Células/tendências , Humanos , Técnicas de Cultura de Órgãos/métodos , Técnicas de Cultura de Órgãos/tendências , Organoides/citologia
15.
J Vis Exp ; (157)2020 03 26.
Artigo em Inglês | MEDLINE | ID: mdl-32281970

RESUMO

Organoids and three-dimensional (3D) cell cultures allow the investigation of complex biological mechanisms and regulations in vitro, which previously was not possible in classical cell culture monolayers. Moreover, monolayer cell cultures are good in vitro model systems but do not represent the complex cellular differentiation processes and functions that rely on 3D structure. This has so far only been possible in animal experiments, which are laborious, time consuming, and hard to assess by optical techniques. Here we describe an assay to quantitatively determine the barrier integrity over time in living small intestinal mouse organoids. To validate our model, we applied interferon gamma (IFN-γ) as a positive control for barrier destruction and organoids derived from IFN-γ receptor 2 knock out mice as a negative control. The assay allowed us to determine the impact of IFN-γ on the intestinal barrier integrity and the IFN-γ induced degradation of the tight junction proteins claudin-2, -7, and -15. This assay could also be used to investigate the impact of chemical compounds, proteins, toxins, bacteria, or patient-derived probes on the intestinal barrier integrity.


Assuntos
Intestinos/fisiologia , Organoides/fisiologia , Animais , Humanos , Processamento de Imagem Assistida por Computador , Camundongos , Modelos Biológicos , Permeabilidade
16.
J Biomed Sci ; 27(1): 32, 2020 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-32035490

RESUMO

BACKGROUND: Fallopian tube epithelial cells (FTEC) were thought to be the origin of high-grade serous ovarian carcinoma (HGSOC). Knowledge of the stemness or initiating characteristics of FTEC is insufficient. Previously, we have characterized the stemness cell marker of FTEC, this study aims to further characterize the clonogenicity and spheroid features of FTEC. METHODS: We successfully derived FTECs from the epithelial layer of the human fallopian tubes. We examined the morphology, proliferation rate, doubling time, and clonal growth of them. At passage 3, the sphere formations on gelatin-coated culture, suspension culture, and matrigel culture were observed, and the expression of LGR5, SSEA3, SSEA4, and other stemness markers was examined. Furthermore, tissue-reconstituted organoids from coculture of FTEC, fallopian stromal cells (FTMSC) and endothelial cells (HUVEC) were examined. RESULTS: FTEC exhibited cuboidal cell morphology and maintained at a constant proliferation rate for up to nine passages (P9). FTEC could proliferate from a single cell with a clonogenic efficiency of 4%. Flow cytometry revealed expressions of normal stem cell markers (SSEA3, SSEA4, and LGR5) and cancer stem cell markers (CD24, CD44, CD117, ROR1, and CD133). FTEC formed spheres and colonies when cultured on low attach dish. In the presence of Matrigel, the stemness and colony formation activity were much enhanced. In co-culturing with FTMSC and HUVEC, FTEC could form organoids that could be blocked by Wnt inhibitor DKK1. Expressions of LGR5 and FOXJ1 expression were also decreased by adding DKK1. CONCLUSION: We demonstrated abundantly presence of stem cells in human FTECs which are efficient in forming colonies, spheres and organoids, relying on Wnt signaling. We also reported for the first time the generation of organoid from reconstitutied cell lineages in the tissue. This may provide a new model for studying the regneration and malignant transformation of the tubal epithelium.


Assuntos
Autorrenovação Celular/fisiologia , Células Epiteliais/fisiologia , Tubas Uterinas/fisiologia , Expressão Gênica , Organoides/fisiologia , Proteínas Wnt/genética , Feminino , Marcadores Genéticos , Humanos , Proteínas Wnt/metabolismo
17.
J Vis Exp ; (155)2020 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-32065155

RESUMO

The development of advanced tumor models has long been encouraged because current cancer models have shown limitations such as lack of three-dimensional (3D) tumor architecture and low relevance to human cancer. Researchers have recently developed a 3D in vitro cancer model referred to as tumor organoids that can mimic the characteristics of a native tumor in a culture dish. Here, experimental procedures are described in detail for the establishment of bladder tumor organoids from a carcinogen-induced murine bladder tumor, including culture, passage, and maintenance of the resulting 3D tumor organoids in vitro. In addition, protocols to manipulate the established bladder tumor organoid lines for genetic engineering using lentivirus-mediated transduction are described, including optimized conditions for the efficient introduction of new genetic elements into tumor organoids. Finally, the procedure for orthotopic transplantation of bladder tumor organoids into the wall of the murine bladder for further analysis is laid out. The methods described in this article can facilitate the establishment of an in vitro model for bladder cancer for the development of better therapeutic options.


Assuntos
Organoides/transplante , Neoplasias da Bexiga Urinária/terapia , Animais , Modelos Animais de Doenças , Camundongos , Organoides/fisiologia
19.
Annu Rev Pathol ; 15: 211-234, 2020 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-31550983

RESUMO

Organoids are in vitro-cultured three-dimensional structures that recapitulate key aspects of in vivo organs. They can be established from pluripotent stem cells and from adult stem cells, the latter being the subject of this review. Organoids derived from adult stem cells exploit the tissue regeneration process that is driven by these cells, and they can be established directly from the healthy or diseased epithelium of many organs. Organoids are amenable to any experimental approach that has been developed for cell lines. Applications in experimental biology involve the modeling of tissue physiology and disease, including malignant, hereditary, and infectious diseases. Biobanks of patient-derived tumor organoids are used in drug development research, and they hold promise for developing personalized and regenerative medicine. In this review, we discuss the applications of adult stem cell-derived organoids in the laboratory and the clinic.


Assuntos
Biologia Celular , Terapia Baseada em Transplante de Células e Tecidos/métodos , Organoides/citologia , Organoides/fisiologia , Adulto , Células-Tronco Adultas/fisiologia , Biologia Celular/tendências , Terapia Baseada em Transplante de Células e Tecidos/tendências , Doença/etiologia , Humanos , Células-Tronco Pluripotentes/fisiologia , Medicina Regenerativa/métodos
20.
J Vis Exp ; (153)2019 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-31789309

RESUMO

At the end of the suckling period, many mammalian species undergo major changes in the intestinal epithelium that are associated with the capability to digest solid food. This process is termed suckling-to-weaning transition and results in the replacement of neonatal epithelium with adult epithelium which goes hand in hand with metabolic and morphological adjustments. These complex developmental changes are the result of a genetic program that is intrinsic to the intestinal epithelial cells but can, to some extent, be modulated by extrinsic factors. Prolonged culture of mouse primary intestinal epithelial cells from late fetal period, recapitulates suckling-to-weaning transition in vitro. Here, we describe a detailed protocol for mouse fetal intestinal organoid culture best suited to model this process in vitro. We describe several useful assays designed to monitor the change of intestinal functions associated with suckling-to-weaning transition over time. Additionally, we include an example of an extrinsic factor that is capable to affect suckling-to-weaning transition in vivo, as a representation of modulating the timing of suckling-to-weaning transition in vitro. This in vitro approach can be used to study molecular mechanisms of the suckling-to-weaning transition as well as modulators of this process. Importantly, with respect to animal ethics in research, replacing in vivo models by this in vitro model contributes to refinement of animal experiments and possibly to a reduction in the use of animals to study gut maturation processes.


Assuntos
Animais Lactentes/fisiologia , Desenvolvimento Fetal/fisiologia , Mucosa Intestinal/embriologia , Mucosa Intestinal/fisiologia , Organoides/embriologia , Organoides/fisiologia , Animais , Células Cultivadas , Células Epiteliais/fisiologia , Mucosa Intestinal/citologia , Camundongos , Técnicas de Cultura de Órgãos , Organoides/citologia , Desmame
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