Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes

Background & Aims: Hepatocyte-like cells (HLCs), differentiated from pluripotent stem cells by the use of soluble factors, can model human liver function and toxicity. However, at present HLC maturity and whether any deficit represents a true fetal state or aberrant differentiation is unclear and compounded by comparison to potentially deteriorated adult hepatocytes. Therefore, we generated HLCs from multiple lineages, using two different protocols, for direct comparison with fresh fetal and adult hepatocytes. Methods: Protocols were developed for robust differentiation. Multiple transcript, protein and functional analyses compared HLCs to fresh human fetal and adult hepatocytes. Results: HLCs were comparable to those of other laboratories by multiple parameters. Transcriptional changes during differentiation mimicked human embryogenesis and showed more similarity to pericentral than periportal hepatocytes. Unbiased proteomics demonstrated greater proximity to liver than 30 other human organs or tissues. However, by comparison to fresh material, HLC maturity was proven by transcript, protein and function to be fetal-like and short of the adult phenotype. The expression of 81% phase 1 enzymes in HLCs was significantly upregulated and half were statistically not different from fetal hepatocytes. HLCs secreted albumin and metabolized testosterone (CYP3A) and dextrorphan (CYP2D6) like fetal hepatocytes. In seven bespoke tests, devised by principal components analysis to distinguish fetal from adult hepatocytes, HLCs from two different source laboratories consistently demonstrated fetal characteristics. Conclusions: HLCs from different sources are broadly comparable with unbiased proteomic evidence for faithful differentiation down the liver lineage. This current phenotype mimics human fetal rather than adult hepatocytes

Théâtre royal de l’Opéra-Comique.

Transcript of THÉÂTRE DE L'OPÉRA-COMIQUE by M.A. Delaforest, appearing in GAZETTE DE FRANCE, 16 août 1822

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[...] Bibliothèque Historique Et Militaire (-) [...] (Tome Premier) ([3]) Einband ( - ) Vortitelblatt ([1]) Titelseite ([3]) Essai Sur La Tactique DesGrecs ([5]) C'est la première fois que l'on essaie de réunir dans une ... ([7]) Chapitre Ier. Principes de l'art militaire consacrés par Homère. - État de l'art depuis la guerre de Troie jusqu' à Cyrus ... ([9]) Chapitre II. Invasion de Darius; bataille de Marathon. - Invasion de Xerxès; combat des Thermopyles (15) Chapitre III. Combat de Salamine. - Xerxès retourne en Asie; Mardonius continue la guerre ... (21) Chapitre IV. Organisation de la Phalange (26) Chapitre V. De la Cavalerie dans ses rapports avec la Phalange (30) Chapitre VI. Mouvemens de la Phalange. - Résumé de ses élémens divers (33) Chapitre VII. Guerre du Péloponnèse. - Première bataille de Mantinée. - Retraite des Dix-Mille (39) Chapitre VIII. Bataille de Leuctres. - Deuxième bataille de Mantinée (43) Chapitre IX. De Philippe et d' Alexandre. - Bataille de Chéronée. - Passage du Granique (47) Chapitre X. Siége d'Halicarnasse. - Darius se décide à porter la guerre en Macédoine. - Memnon de Rhodes meurt au moment d'exécuter ce projet. ... (54) Chapitre XI. Prise de Tyr et de Gaza. - Progrés faits dans la Poliorcetique de ces siéges mémorables. - Fondation d' Alexandrie (59) Chapitre XII. Bataille d'Arbelle. - Passage de l'Hydaspe (63) Chapitre XIII. État de l'art les successeurs d' Alexandre. - De Pyrrhus. - De Philopœmen, surnommé le dernier des Grecs ... (73) Chapitre XIV. Constitution militaire de la Grèce (81) Thucydide (87) Titelseite (89) L'un des plus beaunx caractères de la littérature ancienne, de la littérature grecque en particulier ... (91) Livre Premier (93) Livre Second (148) Livre Troisième (193) Livre Quatrième (238) Livre Cinquième (291) Livre Sixième (330) Livre Septième (373) Livre Huitième (409) Xenophon ([455]) Titelseite ([457]) La traduction que nous réimprimons est faite par un homme également distingué dans la littérature et dans ... ([459]) Livre Premier ([460]) Livre Deuxième (481) Livre Troisième (497) Livre Quatrième (514) Livre Cinquième (534) Livre Sixième (555) Livre Septième (573) Arrien (769) Titelseite (771) Préface D'Arrien (773) Livre Premier (775) Livre Second (797) Livre Troisiéme (814) Livre Quatriéme (833) Livre Cinquiéme (851) Livre Sixiéme (867) Livre Septième (882) Table Des Matières Contenues Dans Le Tome Premier (901) Note A Consulter Pour Le Placement Des Cartes Et Des Plans (903) Errata (904

HNF4A Regulates the Formation of Hepatic Progenitor Cells from Human iPSC-Derived Endoderm by Facilitating Efficient Recruitment of RNA Pol II

Elucidating the molecular basis of cell differentiation will advance our understanding of organ development and disease. We have previously established a protocol that efficiently produces cells with hepatocyte characteristics from human induced pluripotent stem cells. We previously used this cell differentiation model to identify the transcription factor hepatocyte nuclear factor 4 α (HNF4A) as being essential during the transition of the endoderm to a hepatic fate. Here, we sought to define the molecular mechanisms through which HNF4A controls this process. By combining HNF4A chromatin immunoprecipitation (ChIP) followed by high-throughput DNA sequencing (ChIP-seq) analyses at the onset of hepatic progenitor cell formation with transcriptome data collected during early stages of differentiation, we identified genes whose expression is directly dependent upon HNF4A. By examining the dynamic changes that occur at the promoters of these HNF4A targets we reveal that HNF4A is essential for recruitment of RNA polymerase (RNA pol) II to genes that are characteristically expressed as the hepatic progenitors differentiate from the endoderm