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2.
Cells ; 12(15)2023 08 01.
Artículo en Inglés | MEDLINE | ID: mdl-37566061

RESUMEN

Human pluripotent stem cells (hPSCs) are capable of unlimited proliferation and can undergo differentiation to give rise to cells and tissues of the three primary germ layers. While directing lineage selection of hPSCs has been an active area of research, improving the efficiency of differentiation remains an important objective. In this study, we describe a two-compartment microfluidic device for co-cultivation of adult human hepatocytes and stem cells. Both cell types were cultured in a 3D or spheroid format. Adult hepatocytes remained highly functional in the microfluidic device over the course of 4 weeks and served as a source of instructive paracrine cues to drive hepatic differentiation of stem cells cultured in the neighboring compartment. The differentiation of stem cells was more pronounced in microfluidic co-cultures compared to a standard hepatic differentiation protocol. In addition to improving stem cell differentiation outcomes, the microfluidic co-culture system described here may be used for parsing signals and mechanisms controlling hepatic cell fate.


Asunto(s)
Microfluídica , Células Madre Pluripotentes , Humanos , Técnicas de Cocultivo , Microfluídica/métodos , Hepatocitos/metabolismo , Diferenciación Celular
3.
Sci Rep ; 11(1): 7177, 2021 03 30.
Artículo en Inglés | MEDLINE | ID: mdl-33785778

RESUMEN

Cellular therapies based on human pluripotent stem cells (hPSCs) offer considerable promise for treating numerous diseases including diabetes and end stage liver failure. Stem cell spheroids may be cultured in stirred bioreactors to scale up cell production to cell numbers relevant for use in humans. Despite significant progress in bioreactor culture of stem cells, areas for improvement remain. In this study, we demonstrate that microfluidic encapsulation of hPSCs and formation of spheroids. A co-axial droplet microfluidic device was used to fabricate 400 µm diameter capsules with a poly(ethylene glycol) hydrogel shell and an aqueous core. Spheroid formation was demonstrated for three hPSC lines to highlight broad utility of this encapsulation technology. In-capsule differentiation of stem cell spheroids into pancreatic ß-cells in suspension culture was also demonstrated.


Asunto(s)
Técnicas de Cultivo de Célula/métodos , Células Madre Pluripotentes/fisiología , Esferoides Celulares/fisiología , Reactores Biológicos , Cápsulas/química , Técnicas de Cultivo de Célula/instrumentación , Diferenciación Celular , Línea Celular , Supervivencia Celular , Trasplante de Células/métodos , Diabetes Mellitus/terapia , Enfermedad Hepática en Estado Terminal/terapia , Humanos , Hidrogeles/química , Células Secretoras de Insulina/fisiología , Técnicas Analíticas Microfluídicas/instrumentación , Células Madre Pluripotentes/trasplante , Polietilenglicoles/química
4.
Nat Commun ; 11(1): 2241, 2020 05 07.
Artículo en Inglés | MEDLINE | ID: mdl-32382023

RESUMEN

The generation of pancreatic cell types from renewable cell sources holds promise for cell replacement therapies for diabetes. Although most effort has focused on generating pancreatic beta cells, considerable evidence indicates that glucagon secreting alpha cells are critically involved in disease progression and proper glucose control. Here we report on the generation of stem cell-derived human pancreatic alpha (SC-alpha) cells from pluripotent stem cells via a transient pre-alpha cell intermediate. These pre-alpha cells exhibit a transcriptional profile similar to mature alpha cells and although they produce proinsulin protein, they do not secrete significant amounts of processed insulin. Compound screening identified a protein kinase c activator that promotes maturation of pre-alpha cells into SC-alpha cells. The resulting SC-alpha cells do not express insulin, share an ultrastructure similar to cadaveric alpha cells, express and secrete glucagon in response to glucose and some glucagon secretagogues, and elevate blood glucose upon transplantation in mice.


Asunto(s)
Técnicas de Cultivo de Célula/métodos , Células Secretoras de Glucagón/citología , Células Secretoras de Insulina/efectos de los fármacos , Células Madre Pluripotentes/citología , Western Blotting , Diferenciación Celular/fisiología , Línea Celular , Electrofisiología , Técnica del Anticuerpo Fluorescente , Humanos , Páncreas/citología
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