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1.
Cells ; 10(12)2021 12 07.
Artigo em Inglês | MEDLINE | ID: mdl-34943947

RESUMO

Intravital microscopy and other direct-imaging techniques have allowed for a characterisation of leukocyte migration that has revolutionised the field of immunology, resulting in an unprecedented understanding of the mechanisms of immune response and adaptive immunity. However, there is an assumption within the field that modern imaging techniques permit imaging parameters where the resulting cell track accurately captures a cell's motion. This notion is almost entirely untested, and the relationship between what could be observed at a given scale and the underlying cell behaviour is undefined. Insufficient spatial and temporal resolutions within migration assays can result in misrepresentation of important physiologic processes or cause subtle changes in critical cell behaviour to be missed. In this review, we contextualise how scale can affect the perceived migratory behaviour of cells, summarise the limited approaches to mitigate this effect, and establish the need for a widely implemented framework to account for scale and correct observations of cell motion. We then extend the concept of scale to new approaches that seek to bridge the current "black box" between single-cell behaviour and systemic response.


Assuntos
Movimento Celular/fisiologia , Rastreamento de Células/tendências , Leucócitos/fisiologia , Imagem Molecular/tendências , Imunidade Adaptativa/genética , Movimento Celular/genética , Humanos , Imunidade/genética , Leucócitos/ultraestrutura
2.
Front Immunol ; 12: 788891, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-35046946

RESUMO

Blood malignancies provide unique opportunities for longitudinal tracking of disease evolution following therapeutic bottlenecks and for the monitoring of changes in anti-tumor immunity. The expanding development of multi-modal single-cell sequencing technologies affords newer platforms to elucidate the mechanisms underlying these processes at unprecedented resolution. Furthermore, the identification of molecular events that can serve as in-vivo barcodes now facilitate the tracking of the trajectories of malignant and of immune cell populations over time within primary human samples, as these permit unambiguous identification of the clonal lineage of cell populations within heterogeneous phenotypes. Here, we provide an overview of the potential for chromosomal copy number changes, somatic nuclear and mitochondrial DNA mutations, single nucleotide polymorphisms, and T and B cell receptor sequences to serve as personal natural barcodes and review technical implementations in single-cell analysis workflows. Applications of these methodologies include the study of acquired therapeutic resistance and the dissection of donor- and host cellular interactions in the context of allogeneic hematopoietic stem cell transplantation.


Assuntos
Código de Barras de DNA Taxonômico/métodos , Código de Barras de DNA Taxonômico/tendências , Leucemia , Análise de Célula Única/métodos , Análise de Célula Única/tendências , Linhagem da Célula , Rastreamento de Células/métodos , Rastreamento de Células/tendências , Humanos , Imunofenotipagem/métodos , Imunofenotipagem/tendências
3.
Development ; 147(7)2020 04 12.
Artigo em Inglês | MEDLINE | ID: mdl-32280064

RESUMO

Understanding the mechanisms that underlie the generation and regeneration of ß cells is crucial for developing treatments for diabetes. However, traditional research methods, which are based on populations of cells, have limitations for defining the precise processes of ß-cell differentiation and trans-differentiation, and the associated regulatory mechanisms. The recent development of single-cell technologies has enabled re-examination of these processes at a single-cell resolution to uncover intermediate cell states, cellular heterogeneity and molecular trajectories of cell fate specification. Here, we review recent advances in understanding ß-cell generation and regeneration, in vivo and in vitro, from single-cell technologies, which could provide insights for optimization of diabetes therapy strategies.


Assuntos
Diferenciação Celular , Linhagem da Célula , Células Secretoras de Insulina/citologia , Células Secretoras de Insulina/fisiologia , Regeneração/fisiologia , Análise de Célula Única/métodos , Animais , Rastreamento de Células/métodos , Rastreamento de Células/tendências , Humanos , Pâncreas/citologia , Pâncreas/fisiologia , Análise de Célula Única/tendências
4.
Curr Opin Cell Biol ; 63: 114-124, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32058267

RESUMO

Cells rely on a complex network of spatiotemporally regulated signaling activities to effectively transduce information from extracellular cues to intracellular machinery. To probe this activity architecture, researchers have developed an extensive molecular tool kit of fluorescent biosensors and optogenetic actuators capable of monitoring and manipulating various signaling activities with high spatiotemporal precision. The goal of this review is to provide readers with an overview of basic concepts and recent advances in the development and application of genetically encodable biosensors and optogenetic tools for understanding signaling activity.


Assuntos
Clonagem Molecular , Proteínas Luminescentes/genética , Optogenética/métodos , Optogenética/tendências , Transdução de Sinais/genética , Animais , Técnicas Biossensoriais , Rastreamento de Células/métodos , Rastreamento de Células/tendências , Clonagem Molecular/métodos , Humanos , Proteínas Luminescentes/metabolismo , Imagem Molecular/métodos , Imagem Molecular/tendências , Técnicas de Rastreamento Neuroanatômico/métodos , Técnicas de Rastreamento Neuroanatômico/tendências
5.
Theranostics ; 10(4): 1923-1947, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32042345

RESUMO

The continuous or real-time tracking of biological processes using biocompatible contrast agents over a certain period of time is vital for precise diagnosis and treatment, such as monitoring tissue regeneration after stem cell transplantation, understanding the genesis, development, invasion and metastasis of cancer and so on. The rationally designed nanoparticles, including aggregation-induced emission (AIE) dots, inorganic quantum dots (QDs), nanodiamonds, superparamagnetic iron oxide nanoparticles (SPIONs), and semiconducting polymer nanoparticles (SPNs), have been explored to meet this urgent need. In this review, the development and application of these nanoparticle-based cell trackers for a variety of imaging technologies, including fluorescence imaging, photoacoustic imaging, magnetic resonance imaging, magnetic particle imaging, positron emission tomography and single photon emission computing tomography are discussed in detail. Moreover, the further therapeutic treatments using multi-functional trackers endowed with photodynamic and photothermal modalities are also introduced to provide a comprehensive perspective in this promising research field.


Assuntos
Rastreamento de Células/métodos , Nanomedicina Teranóstica/métodos , Animais , Rastreamento de Células/tendências , Meios de Contraste/química , Meios de Contraste/uso terapêutico , Humanos , Imageamento por Ressonância Magnética/métodos , Sondas Moleculares/química , Sondas Moleculares/uso terapêutico , Nanodiamantes/química , Nanopartículas/química , Nanopartículas/uso terapêutico , Imagem Óptica/métodos , Técnicas Fotoacústicas/métodos , Fotoquimioterapia/métodos , Terapia Fototérmica/métodos , Pontos Quânticos/química , Pontos Quânticos/uso terapêutico , Nanomedicina Teranóstica/tendências
6.
Curr Protoc Cell Biol ; 83(1): e88, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31050869

RESUMO

The rapid advancement of live-cell imaging technologies has enabled biologists to generate high-dimensional data to follow biological movement at the microscopic level. Yet, the "perceived" ease of use of modern microscopes has led to challenges whereby sub-optimal data are commonly generated that cannot support quantitative tracking and analysis as a result of various ill-advised decisions made during image acquisition. Even optimally acquired images often require further optimization through digital processing before they can be analyzed. In writing this article, we presume our target audience to be biologists with a foundational understanding of digital image acquisition and processing, who are seeking to understand the essential steps for particle/object tracking experiments. It is with this targeted readership in mind that we review the basic principles of image-processing techniques as well as analysis strategies commonly used for tracking experiments. We conclude this technical survey with a discussion of how movement behavior can be mathematically modeled and described. © 2019 by John Wiley & Sons, Inc.


Assuntos
Rastreamento de Células/métodos , Processamento de Imagem Assistida por Computador/métodos , Modelos Teóricos , Algoritmos , Rastreamento de Células/tendências , Tomada de Decisões , Fluorofotometria , Processamento de Imagem Assistida por Computador/tendências , Razão Sinal-Ruído
8.
Bioconjug Chem ; 28(6): 1581-1597, 2017 06 21.
Artigo em Inglês | MEDLINE | ID: mdl-28485976

RESUMO

Efforts to develop novel cell-based therapies originated with the first bone marrow transplant on a leukemia patient in 1956. Preclinical and clinical examples of cell-based treatment strategies have shown promising results across many disciplines in medicine, with recent advances in immune cell therapies for cancer producing remarkable response rates, even in patients with multiple treatment failures. However, cell-based therapies suffer from inconsistent outcomes, motivating the search for tools that allow monitoring of cell delivery and behavior in vivo. Noninvasive cell imaging techniques, also known as cell tracking, have been developed to address this issue. These tools can allow real-time, quantitative, and long-term monitoring of transplanted cells in the recipient, providing insight on cell migration, distribution, viability, differentiation, and fate, all of which play crucial roles in treatment efficacy. Understanding these parameters allows the optimization of cell choice, delivery route, and dosage for therapy and advances cell-based therapy for specific clinical uses. To date, most cell tracking work has centered on imaging modalities such as MRI, radionuclide imaging, and optical imaging. However, X-ray computed tomography (CT) is an emerging method for cell tracking that has several strengths such as high spatial and temporal resolution, and excellent quantitative capabilities. The advantages of CT for cell tracking are enhanced by its wide availability and cost effectiveness, allowing CT to become one of the most popular clinical imaging modalities and a key asset in disease diagnosis. In this review, we will discuss recent advances in cell tracking methods using X-ray CT in various applications, in addition to predictions on how the field will progress.


Assuntos
Rastreamento de Células/métodos , Neoplasias/diagnóstico por imagem , Rastreamento de Células/tendências , Meios de Contraste , Humanos , Nanopartículas , Neoplasias/patologia , Tomografia Computadorizada por Raios X/métodos , Tomografia Computadorizada por Raios X/tendências
9.
Bioengineered ; 8(4): 316-324, 2017 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-27813700

RESUMO

In contrast to conventional, molecular medicine that focuses on targeting specific pathways, stem cell therapy aims to perturb many related mechanisms in order to derive therapeutic benefit. This emerging modality is inherently complex due to the variety of cell types that can be used, delivery approaches that need to be optimized in order to target the cellular therapeutic to specific sites in vivo, and non-invasive imaging methods that are needed to monitor cell fate. This review highlights advancements in the field, with focus on recent publications that use preclinical animal models for cardiovascular stem cell therapy. It highlights studies where cell adhesion engineering (CAE) has been used to functionalize stem cells to home them to sites of therapy, much like peripheral blood neutrophils. It also describes the current state of molecular imaging approaches that aim to non-invasively track the spatio-temporal pattern of stem cell delivery in living subjects.


Assuntos
Doenças Cardiovasculares/patologia , Doenças Cardiovasculares/terapia , Modelos Animais de Doenças , Imagem Molecular/métodos , Terapia de Alvo Molecular/métodos , Transplante de Células-Tronco/métodos , Células-Tronco/patologia , Animais , Rastreamento de Células/métodos , Rastreamento de Células/tendências , Previsões , Imagem Molecular/tendências , Terapia de Alvo Molecular/tendências , Transplante de Células-Tronco/tendências , Resultado do Tratamento
11.
Discov Med ; 14(75): 97-104, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22935206

RESUMO

Metastasis, the process whereby cancer cells spread from their primary site of origin and grow in adjacent or distant sites, is the primary cause of death in cancer patients. The last 30 years has witnessed significant progress in decreasing cancer mortality rates--largely as a result of improved screening and prevention, practical applications of cancer genomics, and less toxic, more targeted therapies. Despite these improvements, metastasis relentlessly drives mortality. The pervasive mortality from metastasis highlights the shortcomings of traditionally accepted hypotheses on the metastatic process. Historically, metastasis has been described as a unidirectional process, whereby cancer cells leave a primary tumor and seed metastasis in regional lymph nodes or distant sites. This anatomically based hypothesis has dictated much of our medical, and in particular, surgical approach to treating cancers. Alternatively, recent research indicates that metastasis is a multidirectional process whereby cancer cells can seed distant sites as well as the primary tumor itself. The multidirectional pathway of cancer cells, termed "self-seeding," has been corroborated in several experimental and clinical models. This review will evaluate the "self-seeding" hypothesis with attention both to the "seed" (cancer cells) as well as the "soil" (premetastatic niche). Increasingly, the role of the microenvironment surrounding metastases appears essential to the survival of metastatic colonies. The self-seeding model depends not only on the inherent mobility of cancer cells, but also on the supporting non-cancerous cells which enable circulating tumors cells to migrate to and survive in distant sites. The recognition that some of these non-cancerous cells include key components of the immune system has re-ignited the field of immunotherapy in cancer. One particular area of immunotherapy research, tumor entrained neutrophils, will be reviewed in more depth. Ultimately, understanding the dynamic interplay between cancer cells and the metastatic niche offers fertile ground for progress both in the treatment and prevention of metastasis.


Assuntos
Neoplasias da Mama , Carcinoma , Continuidade da Assistência ao Paciente/tendências , Detecção Precoce de Câncer/tendências , Oncologia/tendências , Células-Tronco Neoplásicas/patologia , Animais , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/patologia , Neoplasias da Mama/terapia , Carcinoma/diagnóstico , Carcinoma/patologia , Carcinoma/terapia , Rastreamento de Células/métodos , Rastreamento de Células/tendências , Formação de Conceito , Detecção Precoce de Câncer/métodos , Feminino , Humanos , Oncologia/métodos , Modelos Biológicos , Sementes , Solo/análise
12.
Eur Radiol ; 22(1): 189-204, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21735069

RESUMO

Stem cell therapies hold the great promise and interest for cardiac regeneration among scientists, clinicians and patients. However, advancement and distillation of a standard treatment regimen are not yet finalised. Into this breach step recent developments in the imaging biosciences. Thus far, these technical and protocol refinements have played a critical role not only in the evaluation of the recovery of cardiac function but also in providing important insights into the mechanism of action of stem cells. Molecular imaging, in its many forms, has rapidly become a necessary tool for the validation and optimisation of stem cell engrafting strategies in preclinical studies. These include a suite of radionuclide, magnetic resonance and optical imaging strategies to evaluate non-invasively the fate of transplanted cells. In this review, we highlight the state-of-the-art of the various imaging techniques for cardiac stem cell presenting the strengths and limitations of each approach, with a particular focus on clinical applicability.


Assuntos
Rastreamento de Células/métodos , Cardiopatias/patologia , Células-Tronco Pluripotentes Induzidas/patologia , Imagem Molecular/métodos , Transplante de Células-Tronco/métodos , Rastreamento de Células/tendências , Feminino , Cardiopatias/diagnóstico , Cardiopatias/cirurgia , Humanos , Aumento da Imagem/métodos , Masculino , Imagem Molecular/tendências , Transplante de Células-Tronco/tendências , Resultado do Tratamento
13.
Genesis ; 49(7): 484-7, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21344613

RESUMO

In the mid-1980s live-cell imaging was changed by the introduction of video techniques, allowing new ways to collect and store data. The increased resolution obtained by manipulating video signals, the ability to use time-lapse videocassette recorders to study events that happen over long time intervals, and the introduction of fluorescent probes and sensitive video cameras opened research avenues previously unavailable. The author gives a personal account of this evolution, focusing on cell migration studies at the Marine Biological Laboratory 25 years ago.


Assuntos
Rastreamento de Células , Células/citologia , Animais , Movimento Celular , Rastreamento de Células/história , Rastreamento de Células/instrumentação , Rastreamento de Células/métodos , Rastreamento de Células/tendências , Células/ultraestrutura , Embrião não Mamífero , História do Século XX , História do Século XXI , Humanos , Microscopia de Vídeo
14.
Pediatr Radiol ; 41(2): 199-207, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21127854

RESUMO

Molecular imaging is aimed at the noninvasive in vivo characterization and measurement of processes at a cellular and molecular level with clinical imaging methods. Contrast agents are constructed to target markers that are specific either for certain diseases or for functional states of specialized tissues. Efforts are currently focused mainly on processes involved in angiogenesis, inflammation, and apoptosis. Cell tracking is performed for diagnostic purposes as well as for monitoring of novel cell therapies. Visualization of these processes would provide more precise information about disease expansion as well as treatment response, and could lead to a more individualized therapy for patients. Many attempts have shown promising results in preclinical studies; however, translation into the clinic remains a challenge. This applies especially to paediatrics because of more stringent safety concerns and the low prevalence of individual diseases. The most promising modalities for clinical translation are nuclear medicine methods (positron emission tomography [PET] and single photon emission CT [SPECT]) due to their high sensitivity, which allows concentrations below biological activity. However, special dose consideration is required for any application of ionizing radiation especially in children. While very little has been published on molecular imaging in a paediatric patient population beyond fluorodeoxyglucose (FDG)-PET and metaiodobenzylguanidine (MIBG) tracers, this review will attempt to discuss approaches that we believe have promise for paediatric imaging. These will include agents that already reached clinical trials as well as preclinical developments with high potential for clinical application.


Assuntos
Rastreamento de Células/tendências , Imagem Molecular/tendências , Pediatria/tendências , Radiologia/tendências , Cintilografia/tendências , Criança , Humanos
15.
AJNR Am J Neuroradiol ; 31(9): 1577-83, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20864520

RESUMO

In the foreseeable future, the MI field could greatly assist neuroradiologists. Reporter molecules provide information on specific molecular or cellular events that could not only aid diagnosis but potentially differentiate stages of disorders and treatments. To accomplish this, reporter molecules literally need to pass a barrier, the BBB, which is designed to repel nonessential molecules from the brain. Although this is not a trivial task, several transport systems could be tricked into guiding molecules into the brain. The noninvasive nature in conjunction with a wide availability makes MR imaging particularly suitable for longitudinal neurologic imaging studies. This review explains the principles of MR imaging contrast, delineates different types of reporter molecules, and describes strategies to transport reporters into the brain. It also discusses recent advances in MR imaging hardware, pulse sequences, the development of targeted reporter probes, and future directions of the MR neuroimaging field.


Assuntos
Biomarcadores/análise , Encefalopatias/patologia , Encéfalo/metabolismo , Encéfalo/patologia , Rastreamento de Células/tendências , Imageamento por Ressonância Magnética/tendências , Imagem Molecular/tendências , Previsões , Humanos
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