Automated generation of hiPSC-derived hepatic progeny by cost-efficient compounds.

Fecha de publicación: 24/08/2023 Fecha Ahead of Print: 24/08/2023

Autores de IIS La Fe

Guillem García Llorens

Autor
José Vicente Castell Ripoll

Autor

Participantes ajenos a IIS La Fe

Vanmarcke G
Sai-Hong Chui J
Cooreman A
De Vos K
Cleuren L
Van Rossom R
Izuel Idoype T
Boon R
Kumar Gautam M
Annaert P
Lluis F
Verfaillie CM

Grupos

Hepatología experimental y Trasplante hepático

Investigador Principal

Laia Tolosa Pardo
Unidad Mixta de Investigación en Hepatología Experimental

Investigador Principal

Ramiro Jover Atienza

Abstract

Human pluripotent stem cell (hPSC)-derived hepatocyte-like cells (HLCs) hold great promise for liver disease modeling, drug discovery and drug toxicity screens. Yet, several hurdles still need to be overcome, including among others decrease in the cost of goods to generate HLCs and automation of the differentiation process. We here describe that use of an automated liquid handling system results in highly reproducible HLC differentiation from hPSCs. This enabled us to screen 92 chemicals to replace expensive growth factors at each step of the differentiation protocol to reduce the cost of goods of the differentiation protocol by approximately 79%. In addition, we also evaluated several recombinant extracellular matrices (ECM) to replace Matrigel. We demonstrated that differentiation of hPSCs on Laminin-521 using an optimized small molecule combination resulted in HLCs that were transcriptionally identical to HLCs generated using the growth factor combinations. In addition, the HLCs created using the optimized small molecule combination secreted similar amounts of albumin and urea, and relatively low concentrations of alfa-fetoprotein (AFP), displayed similar CYP3A4 functionality and a similar drug toxicity susceptibility as HLCs generated with growth factor cocktails. The broad applicability of the new differentiation protocol was demonstrated for four different hPSC lines. This allowed the creation of a scalable, xeno-free, and cost-efficient hPSC-derived HLC culture, suitable for high throughput disease modeling and drug screenings, or even for the creation of HLCs for regenerative therapies.

Datos de la publicación

ISSN/ISSNe:: 1066-5099, 1549-4918
Tipo:: Article
Páginas:: 1076-1088
DOI:: 10.1093/stmcls/sxad065
Factor de Impacto:: 1,374 SCImago ℠
Cuartil:: Q1 SCImago ℠

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Métricas

Filiaciones

Vanmarcke G:: Department of Development and Regeneration, Stem Cell Institute, KU Leuven, Leuven 3000, Belgium
Sai-Hong Chui J:: Department of Development and Regeneration, Stem Cell Institute, KU Leuven, Leuven 3000, Belgium
Cooreman A:: Department of Pharmaceutical and Pharmacological Sciences, Vrije Universiteit Brussel, Brussels 1090, Belgium

Drug Delivery and Disposition, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Leuven 3000, Belgium
De Vos K:: Drug Delivery and Disposition, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Leuven 3000, Belgium
Cleuren L:: Department of Development and Regeneration, Stem Cell Institute, KU Leuven, Leuven 3000, Belgium
Van Rossom R:: Department of Development and Regeneration, Stem Cell Institute, KU Leuven, Leuven 3000, Belgium
García-Llorens G:: Instituto de Investigación. Unidad de Hepatología Experimental, Health Research Institute La Fe, and Departamento de Bioquímica y Biología Molecular, Universidad de Valencia, Valencia 46010, Spain, CIBEREHD
Izuel Idoype T:: Department of Development and Regeneration, Stem Cell Institute, KU Leuven, Leuven 3000, Belgium
Boon R:: Department of Development and Regeneration, Stem Cell Institute, KU Leuven, Leuven 3000, Belgium
Kumar Gautam M:: Department of Development and Regeneration, Stem Cell Institute, KU Leuven, Leuven 3000, Belgium
Castell JV:: Instituto de Investigación. Unidad de Hepatología Experimental, Health Research Institute La Fe, and Departamento de Bioquímica y Biología Molecular, Universidad de Valencia, Valencia 46010, Spain, CIBEREHD
Annaert P:: Drug Delivery and Disposition, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Leuven 3000, Belgium
Lluis F:: Department of Development and Regeneration, Stem Cell Institute, KU Leuven, Leuven 3000, Belgium
Verfaillie CM:: Department of Development and Regeneration, Stem Cell Institute, KU Leuven, Leuven 3000, Belgium

Keywords

differentiation; drug screenings; hepatocyte-like cells; human pluripotent stem cells; small molecules

Campos de estudio

Proyectos asociados

DESARROLLO DE UN MODELO HEPATOCELULAR HUMANO DIFERENCIADO PARA ESTUDIOS DE METABOLISMO Y POTENCIAL INDUCTOR DE NUEVOS FARMACOS

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

SAF2003-09353 . 2003

A-CUTE-TOX

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

LSHB-CT-2004-512051-ACUTE TOX . FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2005

DESARROLLO DE NUEVAS ESTRATEGIAS DIAGNOSTICAS BASADAS EN EL ESTUDIO DE LA CAPACIDAD METABOLIZANTE DE LA PIEL Y DEL PAPEL DE LAS CELULAS DENDITRICAS EN LA INDUCCION DE LAS REACCIONES ALERGICAS FRENTE A FARMACOS

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

PARTIC PI05/2290 - FUND IMABIS . INSTITUTO DE SALUD CARLOS III; FUNDACION FIMABIS . 2007

DEVELOPMENT OF A HIGH THROUGHPUT GENOMICS-BASED TEST FOR ASSESSING GENOTOXIC AND CARCINOGENIC PROPERTIES OF CHEMICAL COMPOUNDS IN VITRO

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

CARCINOGENOMICS . COMISION EUROPEA . 2006

LINTOP

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

LSHB-CT-2006-037499 - LINTOP . FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2005

MECANISMOS TRNACRIPCIONALES IMPLICADOS EN LA ETIOLOGIA DEL HIGADO GRASO NO ALCOHOLICO. ESTUDIOS EN UN MODELO CELULAR HUMANO DE ESTEATOSIS Y APLICACION AL TRANSPLANTE DE HEPATOCITOS EN TERAPIA CELULAR.

Investigador Principal: RAMIRO JOVER ATIENZA

PI07/0550 . INSTITUTO DE SALUD CARLOS III . 2007

TU HOSPITAL INVESTIGA PARA TI

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

CCT005-07-00936 . 2007

PROYECTO CAIBER

CAI08/01/0061 . INSTITUTO DE SALUD CARLOS III; FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2009

FORINVITOX

Investigador Principal: MARÍA JOSÉ GÓMEZ-LECHÓN MOLINER

FORINVITOX 1, START UP 2 . REMA; UNIVERSIDAD AUTONOMA BARCELONA; FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2008

RED DE INNOVACION EN TECNOLOGIAS MEDICAS Y SANITARIAS - RED ITEMAS

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

RD09/0077/00156 . INSTITUTO DE SALUD CARLOS III . 2010

CREACION Y MEJORA DE INFRAESTRUCTURAS EN MEDICINA REGENERATIVA

Investigador Principal: FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA

PLE2009-0141 ACI-PLAN E . MINISTERIO DE CIENCIA E INNOVACION; FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2009

METABONOMICA Y CITOMICA COMO NUEVOS PARADIGMAS Y ESTRATEGIAS EXPERIMENTALES PARA EL ESTUDIO DE LOS MECANISMOS DE HEPATOTOXICIDAD DE FARMACOS, PREDICCION DE SU POTENCIAL HEPATOTOXICO Y LA TRANSICION DE HEPATITIS I ATROGENICA A AUTOINMUNE.

Investigador Principal: MARÍA TERESA DONATO MARTÍN

PI10/00923 . INSTITUTO DE SALUD CARLOS III . 2010

INNOVALIV: INNOVATIVE STRATEGIES TO GENERATE HUMAN HEPATOCYTES FOR TREATMENT OF METABOLIC LIVER DISEASES: TOOLS FOR PERSONALIZED CELL THERAPY

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

278152_INNOVALIV_PE_CASTELL_FP7-HEALTH-2011-two-st . COMISION EUROPEA . 2011

PROGRAMA DE TECNICO DE APOYO. MODALIDAD EMPRESAS

Investigador Principal: FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA

PTA2012-8060-E . MINISTERIO DE ECONOMIA Y COMPETITIVIDAD . 2013

SOLICITUD DE CENTRO I+D RAMON Y CAJAL

Investigador Principal: MARÍA ÁNGELES TORMO MAS

RYC-2012-12246 . MINISTERIO DE ECONOMIA Y COMPETITIVIDAD; FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2014