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1.
Eur J Immunol ; 53(3): e2249934, 2023 03.
Artículo en Inglés | MEDLINE | ID: mdl-36645212

RESUMEN

Thymic epithelial cells (TECs) are key effectors of the thymic stroma and are critically required for T-cell development. TECs comprise a diverse set of related but functionally distinct cell types that are scarce and difficult to isolate and handle. This has precluded TEC-based screening assays. We previously described induced thymic epithelial cells (iTECs), an artificial cell type produced in vitro by direct reprogramming, raising the possibility that iTECs might provide the basis for functional screens related to TEC biology. Here, we present an iTEC-based three-stage medium/high-throughput in vitro assay for synthetic polymer mimics of thymic extracellular matrix (ECM). Using this assay, we identified, from a complex library, four polymers that bind iTEC as well as or better than gelatin but do not bind mesenchymal cells. We show that these four polymers also bind and maintain native mouse fetal TECs and native human fetal TECs. Finally, we show that the selected polymers do not interfere with iTEC function or T-cell development. Collectively, our data establish that iTECs can be used to screen for TEC-relevant compounds in at least some medium/high-throughput assays and identify synthetic polymer ECM mimics that can replace gelatin or ECM components in TEC culture protocols.


Asunto(s)
Gelatina , Timo , Ratones , Humanos , Animales , Gelatina/metabolismo , Células Epiteliales/metabolismo , Diferenciación Celular , Matriz Extracelular
2.
Chem Soc Rev ; 51(20): 8584-8611, 2022 Oct 17.
Artículo en Inglés | MEDLINE | ID: mdl-36165090

RESUMEN

Pathogenic microorganisms are considered to a major threat to human health, impinging on multiple sectors including hospitals, dentistry, food storage and packaging, and water contamination. Due to the increasing levels of antimicrobial resistance shown by pathogens, often caused by long-term abuse or overuse of traditional antimicrobial drugs, new approaches and solutions are necessary. In this area, antimicrobial polymers are a viable solution to combat a variety of pathogens in a number of contexts. Indeed, polymers with intrinsic antimicrobial activities have long been an intriguing research area, in part, due to their widespread natural abundance in materials such as chitin, chitosan, carrageen, pectin, and the fact that they can be tethered to surfaces without losing their antimicrobial activities. In addition, since the discovery of the strong antimicrobial activity of some synthetic polymers, much work has focused on revealing the most effective structural elements that give rise to optimal antimicrobial properties. This has often been synthesis targeted, with the generation of either new polymers or the modification of natural antimicrobial polymers with the addition of antimicrobial enhancing modalities such as quaternary ammonium or guanidinium groups. In this review, the growing number of polymers showing intrinsic antimicrobial properties from the past decade are highlighted in terms of synthesis; often based on post-synthesis modification and their utilization. This includes as surface coatings, for example on medical devices, such as intravascular catheters, orthopaedic implants and contact lenses, or directly as antibacterial agents (specifically as eye drops). Surface functionalisation with inherently antimicrobial polymers is highlighted and has been achieved via various techniques, including surface-bound initiators allowing RAFT or ATRP surface-based polymerization, or via physical immobilization such as by layer-by-layer techniques. This article also covers the mechanistic modes of action of intrinsic antimicrobial polymers against bacteria, viruses, or fungi.


Asunto(s)
Compuestos de Amonio , Antiinfecciosos , Quitosano , Antibacterianos/química , Antibacterianos/farmacología , Antiinfecciosos/química , Antiinfecciosos/farmacología , Quitosano/química , Guanidina , Humanos , Soluciones Oftálmicas , Pectinas , Polímeros/química , Polímeros/farmacología , Agua
3.
Chem Soc Rev ; 51(6): 2081-2120, 2022 Mar 21.
Artículo en Inglés | MEDLINE | ID: mdl-35188510

RESUMEN

Proteases are excellent biomarkers for a variety of diseases, offer multiple opportunities for diagnostic applications and are valuable targets for therapy. From a chemistry-based perspective this review discusses and critiques the most recent advances in the field of substrate-based probes for the detection and analysis of proteolytic activity both in vitro and in vivo.


Asunto(s)
Péptido Hidrolasas , Péptidos , Biomarcadores , Péptido Hidrolasas/metabolismo , Péptidos/metabolismo , Proteolisis
4.
Eur Respir J ; 57(3)2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33060152

RESUMEN

Solitary pulmonary nodules (SPNs) are a clinical challenge, given there is no single clinical sign or radiological feature that definitively identifies a benign from a malignant SPN. The early detection of lung cancer has a huge impact on survival outcome. Consequently, there is great interest in the prompt diagnosis, and treatment of malignant SPNs. Current diagnostic pathways involve endobronchial/transthoracic tissue biopsies or radiological surveillance, which can be associated with suboptimal diagnostic yield, healthcare costs and patient anxiety. Cutting-edge technologies are needed to disrupt and improve, existing care pathways. Optical fibre-based techniques, which can be delivered via the working channel of a bronchoscope or via transthoracic needle, may deliver advanced diagnostic capabilities in patients with SPNs. Optical endomicroscopy, an autofluorescence-based imaging technique, demonstrates abnormal alveolar structure in SPNs in vivo Alternative optical fingerprinting approaches, such as time-resolved fluorescence spectroscopy and fluorescence-lifetime imaging microscopy, have shown promise in discriminating lung cancer from surrounding healthy tissue. Whilst fibre-based Raman spectroscopy has enabled real-time characterisation of SPNs in vivo Fibre-based technologies have the potential to enable in situ characterisation and real-time microscopic imaging of SPNs, which could aid immediate treatment decisions in patients with SPNs. This review discusses advances in current imaging modalities for evaluating SPNs, including computed tomography (CT) and positron emission tomography-CT. It explores the emergence of optical fibre-based technologies, and discusses their potential role in patients with SPNs and suspected lung cancer.


Asunto(s)
Neoplasias Pulmonares , Nódulo Pulmonar Solitario , Humanos , Neoplasias Pulmonares/diagnóstico por imagen , Fibras Ópticas , Tomografía Computarizada por Tomografía de Emisión de Positrones , Nódulo Pulmonar Solitario/diagnóstico por imagen , Tomografía Computarizada por Rayos X
5.
Eur J Nucl Med Mol Imaging ; 48(3): 800-807, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-32915268

RESUMEN

PURPOSE: The relentless rise in antimicrobial resistance is a major societal challenge and requires, as part of its solution, a better understanding of bacterial colonization and infection. To facilitate this, we developed a highly efficient no-wash red optical molecular imaging agent that enables the rapid, selective, and specific visualization of Gram-positive bacteria through a bespoke optical fiber-based delivery/imaging endoscopic device. METHODS: We rationally designed a no-wash, red, Gram-positive-specific molecular imaging agent (Merocy-Van) based on vancomycin and an environmental merocyanine dye. We demonstrated the specificity and utility of the imaging agent in escalating in vitro and ex vivo whole human lung models (n = 3), utilizing a bespoke fiber-based delivery and imaging device, coupled to a wide-field, two-color endomicroscopy system. RESULTS: The imaging agent (Merocy-Van) was specific to Gram-positive bacteria and enabled no-wash imaging of S. aureus within the alveolar space of whole ex vivo human lungs within 60 s of delivery into the field-of-view, using the novel imaging/delivery endomicroscopy device. CONCLUSION: This platform enables the rapid and specific detection of Gram-positive bacteria in the human lung.


Asunto(s)
Fibras Ópticas , Staphylococcus aureus , Endoscopios , Bacterias Grampositivas , Humanos , Pulmón/diagnóstico por imagen
6.
Bioorg Med Chem ; 44: 116298, 2021 08 15.
Artículo en Inglés | MEDLINE | ID: mdl-34243043

RESUMEN

Antibody-directed enzyme prodrug therapy (ADEPT) is a powerful concept in which antibody targeting is linked to enzymatic prodrug activation. The work herein describes the first steps in the development of a technology analogous to ADEPT but in which a palladium catalyst is attached of an antibody rather than an enzyme. Antibody-metal conjugates have been used in a variety of contexts including for radiotherapy; however, none of the metals attached to the antibodies have been used for catalytic purposes. This work represents the first example a metal being attached to an antibody for the purposes of carrying a functional catalyst.


Asunto(s)
Anticuerpos/química , Paladio/química , Catálisis , Estructura Molecular
7.
Molecules ; 26(2)2021 Jan 12.
Artículo en Inglés | MEDLINE | ID: mdl-33445514

RESUMEN

The cellular plasma membrane plays a fundamental role in biological processes, including cell growth, signaling and transport. The labelling of the plasma membrane with targeted fluorescent probes offers a convenient and non-invasive way to image the morphological changes and dynamics of a membrane in real-time and, despite many examples of fluorescent plasma membrane probes, a "universal targeting/anchoring moiety" is still required. In this study, a small number of stearic acid-based probes labelled with 6-carboxyfluorescein was designed and fabricated via solid-phase synthesis in which variations in both charge and hydrophobicity were explored. To ease the synthesis process, a gram-scale synthesis of the Fmoc-Lys(6-carboxyfluoresein diacetate)-OH building block was developed, allowing the discovery of optimal probes that carried a positively charged amino group and a stearic acid tail that exhibited intense plasma membrane brightness and robust retention.


Asunto(s)
Membrana Celular/metabolismo , Colorantes Fluorescentes/síntesis química , Técnicas de Síntesis en Fase Sólida , Coloración y Etiquetado , Colorantes Fluorescentes/química , Células HeLa , Humanos
8.
Clin Infect Dis ; 70(12): 2561-2567, 2020 06 10.
Artículo en Inglés | MEDLINE | ID: mdl-31343064

RESUMEN

BACKGROUND: The Global Programme to Eliminate Lymphatic Filariasis (GPELF) was launched in 2000 with the goal of eliminating lymphatic filariasis (LF) as a public health problem by 2020. Despite considerable progress, the current prevalence is around 60% of the 2000 figure, with the deadline looming a year away. Consequently, there is a continued need for investment in both the mass drug administration (MDA) and morbidity management programs, and this paper aims to demonstrate that need by estimating the health and economic burdens of LF prior to MDA programs starting in GPELF areas. METHODS: A previously developed model was used to estimate the numbers of individuals infected and individuals with symptomatic disease, along with the attributable number of disability-adjusted life years (DALYs). The economic burden was calculated by quantifying the costs incurred by the health-care system in managing clinical cases, the patients' out-of-pocket costs, and their productivity costs. RESULTS: Prior to the MDA program, approximately 129 million people were infected with LF, of which 43 million had clinical disease, corresponding to a DALY burden of 5.25 million. The average annual economic burden per chronic case was US $115, the majority of which resulted from productivity costs. The total economic burden of LF was estimated at US $5.8 billion annually. CONCLUSIONS: These results demonstrate the magnitude of the LF burden and highlight the continued need to support the GPELF. Patients with clinical disease bore the majority of the economic burden, but will not benefit much from the current MDA program, which is aimed at reducing transmission. This assessment further highlights the need to scale up morbidity management programs.


Asunto(s)
Filariasis Linfática , Costo de Enfermedad , Filariasis Linfática/tratamiento farmacológico , Filariasis Linfática/epidemiología , Filariasis Linfática/prevención & control , Humanos , Administración Masiva de Medicamentos , Salud Pública , Años de Vida Ajustados por Calidad de Vida
9.
J Am Chem Soc ; 142(52): 21615-21621, 2020 12 30.
Artículo en Inglés | MEDLINE | ID: mdl-33326212

RESUMEN

With the aid of bioorthogonal chemistry, we demonstrate the fabrication of synthetic dendrimers in situ around living cells. Using tetrazine dienophile and aminooxyl/hydrazide aldehyde chemistries, the density of functional groups on the dendrimers exponentially amplified intensities of fluorescent markers in antibody-targeted live cell imaging. This novel "swarming" approach highlights the power of bioorthogonal chemistry and provides a route to non-natural chemical structures on cells, paving the way for the generation of various artificial cellular nanostructures and scaffolds.

10.
Opt Lett ; 45(10): 2716-2719, 2020 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-32412449

RESUMEN

A robust method to selectively attach specific fluorophores onto the individual cores of a multicore fiber is reported in this Letter. The method is based on the use of ultrafast laser pulses to nanostructure the facet of the fiber core, followed by amine functionalization and sensor conjugation. This surface-machining protocol not only enables precise spatial selectivity, but it also facilitates high deposition densities of the sensor moieties. As a proof of concept, the successful deposition of three different fluorophores onto selected cores of a multicore fiber is demonstrated. The protocol was developed to include attachment of a fluorescence-based pH sensor using the ratiometric carboxynapthofluorescein.

11.
Analyst ; 145(3): 975-982, 2020 Feb 03.
Artículo en Inglés | MEDLINE | ID: mdl-31829318

RESUMEN

Proteases are ideal target biomarkers as they have been implicated in many disease states, including steps associated with cancer progression. Electrochemical peptide-based biosensors have attracted much interest in recent years. However, the significantly large size of the electrodes typically used in most of these platforms has led to performance limitations. These could be addressed by the enhancements offered by microelectrodes, such as rapid response times, improved mass transport, higher signal-to-noise and sensitivity, as well as more localised and less invasive measurements. We present the production and characterisation of a miniaturised electrochemical biosensor for the detection of trypsin, based on 25 µm diameter Pt microelectrodes (rather than the ubiquitous Au electrodes), benchmarked by establishing the equivalent Pt macroelectrode response in terms of quantitative response to the protease, the kinetics of cleavage and the effects of non-specific protein binding and temperature. Interestingly, although there was little difference between Au and Pt macroelectrode response, significant differences were observed between the responses of the Pt macroelectrode and microelectrode systems indicative of increased reproducibility in the microelectrode SAM structure and sensor performance between the electrodes, increased storage stability and a decrease in the cleavage rate at functionalised microelectrodes, which is mitigated by measurement at normal body temperature. Together, these results demonstrate the robustness and sensitivity of the miniaturised sensing platform and its ability to operate within the clinically-relevant concentration ranges of proteases in normal and disease states. These are critical features for its translation into implantable devices.


Asunto(s)
Técnicas Biosensibles/métodos , Péptidos/metabolismo , Platino (Metal)/química , Tripsina/análisis , Técnicas Biosensibles/instrumentación , Técnicas Electroquímicas , Cinética , Microelectrodos , Miniaturización , Péptidos/química , Temperatura , Tripsina/metabolismo
12.
Sensors (Basel) ; 20(21)2020 Oct 27.
Artículo en Inglés | MEDLINE | ID: mdl-33121191

RESUMEN

Numerous optodes, with fluorophores as the chemical sensing element and optical fibres for light delivery and collection, have been fabricated for minimally invasive endoscopic measurements of key physiological parameters such as pH. These flexible miniaturised optodes have typically attempted to maximize signal-to-noise through the application of high concentrations of fluorophores. We show that high-density attachment of carboxyfluorescein onto silica microspheres, the sensing elements, results in fluorescence energy transfer, manifesting as reduced fluorescence intensity and lifetime in addition to spectral changes. We demonstrate that the change in fluorescence intensity of carboxyfluorescein with pH in this "high-density" regime is opposite to that normally observed, with complex variations in fluorescent lifetime across the emission spectra of coupled fluorophores. Improved understanding of such highly loaded sensor beads is important because it leads to large increases in photostability and will aid the development of compact fibre probes, suitable for clinical applications. The time-resolved spectral measurement techniques presented here can be further applied to similar studies of other optodes.

13.
Molecules ; 25(22)2020 Nov 10.
Artículo en Inglés | MEDLINE | ID: mdl-33182751

RESUMEN

Photodynamic inactivation of microorganisms has gained substantial attention due to its unique mode of action, in which pathogens are unable to generate resistance, and due to the fact that it can be applied in a minimally invasive manner. In photodynamic therapy (PDT), a non-toxic photosensitizer (PS) is activated by a specific wavelength of light and generates highly cytotoxic reactive oxygen species (ROS) such as superoxide (O2-, type-I mechanism) or singlet oxygen (1O2*, type-II mechanism). Although it offers many advantages over conventional treatment methods, ROS-mediated microbial killing is often faced with the issues of accessibility, poor selectivity and off-target damage. Thus, several strategies have been employed to develop target-specific antimicrobial PDT (aPDT). This includes conjugation of known PS building-blocks to either non-specific cationic moieties or target-specific antibiotics and antimicrobial peptides, or combining them with targeting nanomaterials. In this review, we summarise these general strategies and related challenges, and highlight recent developments in targeted aPDT.


Asunto(s)
Antiinfecciosos/farmacología , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/síntesis química , Fármacos Fotosensibilizantes/farmacología , Animales , Humanos , Concentración de Iones de Hidrógeno , Liposomas/química , Ratones , Micelas , Oligosacáridos/química , Péptidos/química , Polímeros/química , Oxígeno Singlete/química , Electricidad Estática , Superóxidos
14.
Chembiochem ; 20(7): 872-876, 2019 04 01.
Artículo en Inglés | MEDLINE | ID: mdl-30394615

RESUMEN

Traditionally, prodrug activation has been limited to enzymatic triggers or gross physiological aberrations, such as pH, that offer low selectivity and control over dosage. In recent years, the field of prodrug activation chemistry has been transformed by the use of bioorthogonal reactions that can be carried out under biological conditions at sub-millimolar concentrations, with the tetrazine-mediated inverse electron demand Diels-Alder reaction amongst the most recognised. Their high reaction rates, chemoselectivity and excellent biocompatibility make tetrazines ideal small molecules for activating prodrugs. Recently the tetrazine moiety has been used as a prodrug for a pyridazine thus broadening the scope of prodrug systems. This article discusses the concept of using tetrazines as small-molecule activators for prodrugs, and provides an overview of tetrazine-based prodrug systems, with a particular focus on the recently reported prodrug-prodrug activation strategy.


Asunto(s)
Compuestos Heterocíclicos con 1 Anillo/química , Profármacos/química , Línea Celular Tumoral , Química Clic , Reacción de Cicloadición , Humanos , Piridazinas/síntesis química
15.
Org Biomol Chem ; 17(22): 5533-5537, 2019 06 05.
Artículo en Inglés | MEDLINE | ID: mdl-31090781

RESUMEN

Taking inspiration from the assembly of so-called peptoids (N-alkylglycine oligomers) we present a new synthetic methodology whereby N-heterocyclic carbene (NHC) based Pd ligands were assembled using a sub-monomer approach and loaded with Pd via solid-phase synthesis. This allowed the rapid generation a library of NHC-palladium catalysts that were readily functionalised to allow bioconjugation. These catalysts were able to rapidly activate a caged fluorophore and 'switch-on' an anticancer prodrug in 3D cell culture.


Asunto(s)
Materiales Biocompatibles/síntesis química , Compuestos Heterocíclicos/síntesis química , Metano/análogos & derivados , Paladio/química , Técnicas de Síntesis en Fase Sólida , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Catálisis , Supervivencia Celular/efectos de los fármacos , Compuestos Heterocíclicos/química , Compuestos Heterocíclicos/farmacología , Humanos , Ligandos , Células MCF-7 , Metano/síntesis química , Metano/química , Metano/farmacología , Estructura Molecular
16.
Angew Chem Int Ed Engl ; 58(40): 14189-14192, 2019 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-31397963

RESUMEN

A ruthenium-based mitochondrial-targeting photosensitiser that undergoes efficient cell uptake, enables the rapid catalytic conversion of PtIV prodrugs into their active PtII counterparts, and drives the generation of singlet oxygen was designed. This dual mode of action drives two orthogonal cancer-cell killing mechanisms with temporal and spatial control. The designed photosensitiser was shown to elicit cell death of a panel of cancer cell lines including those showing oxaliplatin-resistance.


Asunto(s)
Antineoplásicos/farmacología , Compuestos Organoplatinos/farmacología , Fármacos Fotosensibilizantes/farmacología , Profármacos/farmacología , Oxígeno Singlete/metabolismo , Antineoplásicos/síntesis química , Antineoplásicos/química , Catálisis , Muerte Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Estructura Molecular , Compuestos Organoplatinos/síntesis química , Compuestos Organoplatinos/química , Procesos Fotoquímicos , Fotoquimioterapia , Fármacos Fotosensibilizantes/síntesis química , Fármacos Fotosensibilizantes/química , Profármacos/síntesis química , Profármacos/química , Oxígeno Singlete/química
17.
Biomacromolecules ; 19(7): 2721-2730, 2018 07 09.
Artículo en Inglés | MEDLINE | ID: mdl-29689166

RESUMEN

The combination of controlled living polymerization in association with rapid and highly efficient macromolecule conjugation strategies provides a powerful tool for the synthesis of novel polymeric materials. Here functional block copolymers were rapidly and quantitatively conjugated using an efficient reaction between polymers containing a phenolic group and the 4-phenyl-3 H-1,2,4-triazole-3,5(4 H)-dione (PTAD) moiety and used to generate nanoparticles that encapsulated drugs. pH responsive amphiphilic block copolymers, which self-assemble into nanoparticles, were fabricated using our novel polymer conjugation strategy with the resulting system designed to promote drug release within the acidic milieu of the cancer microenvironment. The conjugation strategy also enabled the direct tagging of the nanoparticles with a range of fluorophores, targeting assets, or both with cargo release demonstrated in cancer cells.


Asunto(s)
Antineoplásicos/administración & dosificación , Nanoconjugados/química , Técnicas de Química Sintética/métodos , Células HeLa , Humanos , Polimerizacion , Tensoactivos/química , Triazoles/química
18.
Org Biomol Chem ; 16(43): 8056-8063, 2018 11 07.
Artículo en Inglés | MEDLINE | ID: mdl-30175355

RESUMEN

Optical biosensing based on the activation of fluorescent reporters offers a powerful methodology for the real-time molecular interrogation of pathology. Here we report a first-in-class, bimodal fluorescent reporter strategy for the simultaneous and highly specific detection of two independent proteases (thrombin and matrix metalloproteases (MMPs)) pivotal in the fibroproliferative process surrounding lung cancer, based on a dual, multiplexing, peptide FRET system. This sophisticated synthetic smartprobe, with a molecular weight of 6 kDa, contains two independent fluorophores and quenchers that generate photonic signatures at two specific wavelengths upon activation by target enzymes within human lung cancer tissue.


Asunto(s)
Técnicas Biosensibles/métodos , Neoplasias Pulmonares/metabolismo , Proteolisis , Colorantes Fluorescentes/química , Colorantes Fluorescentes/metabolismo , Humanos , Neoplasias Pulmonares/patología , Metaloproteinasas de la Matriz/metabolismo , Neutrófilos/metabolismo , Placa Aterosclerótica/metabolismo
19.
Bioorg Med Chem ; 26(10): 2816-2826, 2018 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-29042225

RESUMEN

Optical medical imaging is a rapidly growing area of research and development that offers a multitude of healthcare solutions both diagnostically and therapeutically. In this review, some of the most recently described peptide-based optical probes are reviewed with a special emphasis on their in vivo use and potential application in a clinical setting.


Asunto(s)
Colorantes Fluorescentes/química , Imagen Óptica/métodos , Péptidos/química , Marcadores de Afinidad/química , Animales , Humanos , Microscopía Fluorescente/métodos
20.
Arch Toxicol ; 92(10): 3117-3129, 2018 10.
Artículo en Inglés | MEDLINE | ID: mdl-30155720

RESUMEN

Liver disease is an escalating global health issue. While liver transplantation is an effective mode of therapy, patient mortality has increased due to the shortage of donor organs. Developing renewable sources of human liver tissue is therefore attractive. Pluripotent stem cell-derived liver tissue represents a potential alternative to cadaver derived hepatocytes and whole organ transplant. At present, two-dimensional differentiation procedures deliver tissue lacking certain functions and long-term stability. Efforts to overcome these limiting factors have led to the building of three-dimensional (3D) cellular aggregates. Although enabling for the field, their widespread application is limited due to their reliance on variable biological components. Our studies focused on the development of 3D liver tissue under defined conditions. In vitro generated 3D tissues exhibited stable phenotype for over 1 year in culture, providing an attractive resource for long-term in vitro studies. Moreover, 3D derived tissue provided critical liver support in two animal models, including immunocompetent recipients. Therefore, we believe that our study provides stable human tissue to better model liver biology 'in the dish', and in the future may permit the support of compromised liver function in humans.


Asunto(s)
Trasplante de Hígado/métodos , Hígado/citología , Células Madre Pluripotentes/citología , Ingeniería de Tejidos/métodos , Animales , Técnicas de Cultivo de Célula , Diferenciación Celular , Endodermo/citología , Femenino , Hepatectomía , Humanos , Hígado/fisiología , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Células Madre Pluripotentes/fisiología , Esferoides Celulares/citología , Factores de Tiempo , Andamios del Tejido
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