Agradecemos a nuestros colegas en RIKEN-BSI por su amable apoyo y consejos durante este fructífero trabajo. Me gustaría expresar mi sincero agradecimiento al profesor Shigemi Mori, profesor G frontera Juh Qué, los Dres. Kiyoji Matsuyama, Katalin AK Kesi y rp d Dobolyi que me aconsejó que método de estudio de microdiálisis en el curso de postgrado en la Universidad de Postgrado para Estudios Avanzados, Japón. Finalmente, agradecemos al Sr. Chris Sarnecky y el Dr. Akitsugu Takasu por su apoyo en la grabación de vídeo.

Paso 1. Preparación de la implantación de la sonda <ol><li> Anestesia (no se muestra en el video) Anestesiar a cada ave con una inyección intraperitoneal de ketamina (12,5 mg / kg de peso corporal) y xilazina (25 mg / kg) de la mezcla para producir un nivel constante de anestesia quirúrgica. </li><li> Fijar la cabeza a un marco estereotáxico El uso de xilocaína (TM) jalea para reducir los dolores locales de bares de presión del oído &quot;. La cabeza de pájaro se fija en un aparato estereotáxico que consiste en bares y oído el titular de pico que tiene la cabeza en un ángulo de 45 °. Durante la cirugía estereotáxica para la implantación, calentar el cuerpo del ave por plataforma quirúrgica con control térmico (temperatura = 34 ~ 36 ° C). </li><li> Desinfectar el área quirúrgica Desinfectar el campo quirúrgico con Isodine (TM) antes de la cirugía. </li></ol> Paso 2. Posicionamiento estereotáxico <ol><li> Referencia punto de alineación Alinear la sonda ficticia (vidrio micropipeta) a la Lambda (Y punto) y la marca Y-punto con la pipeta llena de tinte. Restantes Dextran-TMR del estudio de seguimiento última se utilizó en el video. </li><li> Incisión de la piel Quite la piel y exponer la superficie del cráneo. Luego, limpie la superficie del cráneo con hisopos de algodón. </li><li> Colocación de la cánula dummy (en sentido horizontal) Mover la cánula maniquí al sitio de destino (AP eje y la coordinación lateral) </li><li> Craneotomía pequeña y la incisión de la duramadre. Use unas pinzas finas para craneotomía y una fina aguja de inyección (30 G) para las incisiones de la duramadre. Luego, limpie la superficie del cráneo. </li><li> Posicionamiento final (implantación) de la cánula maniquí Localizar la punta de la cánula guía en el 0,30 mm por encima de la sonda de microdiálisis intencionados región de destino. Por último, limpiar la superficie del cráneo, de nuevo, porque una superficie limpia y seca se requiere para la unión dura. </li></ol> Paso 3. El tratamiento previo de la superficie craneal (El procedimiento similar para la &quot;restauración de obturación directa&quot; del producto dental se emplea). <ol><li> Primer tratamiento <ol><li> Mix PRIMAR A con B líquido en un plato. </li><li> Aplique la mezcla en la superficie del cráneo con un pequeño cepillo. </li><li> Dejar durante 30 segundos. </li><li> Seca con corriente de aire templado. (No se lave.) </li></ol></li><li> Aplicación de agente de unión <ol><li> Aplicar el adhesivo sobre la superficie del cráneo con un pequeño cepillo. </li><li> Aplicar una corriente suave de aire. (No se lave.) </li><li> Fotocurado durante 20 segundos. </li></ol></li></ol> Paso 4. Montaje de resina compuesta <ol><li> Montaje de resina con condensador plano Si un condensador normal no está disponible, una espátula de micro-se puede utilizar en su lugar. </li><li> Fotopolimerización </li><li> Incremental de montaje Incremental de montaje se recomienda para asegurar la reacción completa de la resina. Para el montaje incremental, aplique adhesivo antes de cada montaje. </li><li> Molde de la resina compuesta y la luz de curado de endurecimiento final. </li><li> Final de la cirugía </li></ol> Limpie el área quirúrgica, se aplican los antibióticos (pomada de gentamicina), y retire el animal de la llama estereotáxica. Entonces, mantener al animal en una cámara de termo-control (temperatura = 33-35 ° C) para la recuperación postanestésica. Como se muestra en el video, no se observó un comportamiento anormal en cualquiera de los animales al día siguiente después de la cirugía.

<ol>
	<li>Adell, A., Artigas, F., Boulton, A., Baker, G., Bateson, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+Brain+microdialysis:+Principles+and+applications.">In vivo Brain microdialysis: Principles and applications.</a> In Vivo Neuromethods: Neuromethods. 32, 325-357 (1998).</li><li>Kendrick, K., Robinson, T., Justice, J., Bateson, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microdialysis+in+large+unrestrained+animals:+neuroendocrine+and+behavioral+studies+of+acetylcholine,+amino+acid,+monoamine+and+neuropeptide+release+in+the+sheep.">Microdialysis in large unrestrained animals: neuroendocrine and behavioral studies of acetylcholine, amino acid, monoamine and neuropeptide release in the sheep.</a> Microdialysis in the Neuroscience. 7, 327-348 (1991).</li><li>Oakley, B., Schafer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Experimental Neurobiology: A laboratory Manual. , (1978).</li><li>Okumura, T., Yamashita, Y., Okanoya, K., Tani, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Function+of+the+sensori-motor+nucleus+NIf+in+generation+of+complex+syntactical+song+in+the+Bengalese+Finch+I.+A+Biological+study.">Function of the sensori-motor nucleus NIf in generation of complex syntactical song in the Bengalese Finch I. A Biological study.</a> Soc. Neurosci Abst. 33, (2007).</li><li>Boulton, A., Baker, G., Robinson, T., Bateson, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relation+of+brain+electrical+activity+to+behavior.">The relation of brain electrical activity to behavior.</a> In Vivo Neuromethods: Neuromethods. 32, 325-357 (1998).</li><li>Yamashita, Y., Okumura, T., Okanoya, K., Tani, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Function+of+the+sensori-motor+nucleus+NIf+in+generation+of+complex+syntactical+song+in+the+Bengalese+finch.">Function of the sensori-motor nucleus NIf in generation of complex syntactical song in the Bengalese finch.</a> , 101-104 (2007).</li></ol>

Los experimentos se llevaron a cabo de acuerdo con las directrices para la experimentación con animales del cerebro instituto de ciencias, RIKEN (RIKEN-BSI) y de los animales del instituto del comité de ética aprobado.

En comparación con la aplicación de resinas acrílicas convencionales auto curación, la técnica introducida tiene varias ventajas importantes: 1. Contacto directo: Acrílico autocurado se inserta en el hueso por un tiempo pero no se adhiere a los huesos. Por lo tanto, los tornillos de anclaje están obligados a fijar una base de montaje (ver Figura 2a). Por el contrario, la resina de fotopolimerización pueden unirse al hueso directamente, de forma que los tornillos no son necesarios (Figura 2B). Por lo tanto, el t...

1A fotografía muestra a la sonda de microdiálisis (a), una tuerca (b) y una cánula guía (c). En esta foto, la cánula guía se encuentra rodeado por una resina dental que se adhiere al cráneo directamente. Foto B muestra un pájaro durante la perfusión de drogas. En la parte superior de la cabeza, hay una sonda de microdiálisis que está conectado a un tubo de Tefron bien. Las barras de escala = 1 cm de A y 2 cm de B, respectivamente. <img src="http://www.jove.com/files/249_2008_8_26_3/249_Figure-01.png" /> Figura 1 - Las fotografías de una sonda de microdiálisis (A) y un pájaro durante la perfusión de drogas (B)

application of light-cured dental adhesive resin for mounting electrodes or microdialysis probes in chronic experiments

En los experimentos de grabación crónica, auto-curado de resinas de acrílico dental se han utilizado como una base de montaje de los electrodos o sondas de microdiálisis. Dado que estos acrílicos no se unen a los huesos, tornillos han sido utilizados como anclajes. Sin embargo, en los pequeños animales de experimentación, como los pinzones o el ratón, sus cráneos son muy frágiles y no se puede sostener con éxito los anclajes. En este informe, proponemos una nueva aplicación de la luz de curado de resinas dentales base de montaje de los electrodos o sondas de microdiálisis en experimentos crónicos. Este material permite contacto directo con el cráneo. Por lo tanto, los tornillos de anclaje no son necesarias y campo quirúrgico se reduce considerablemente. Las experiencias pasadas muestran que el efecto de la vinculación mantiene más de 2 meses. Ventana de resina convencional de tiempo en que los materiales son flexibles y viables, a pocos minutos. Sin embargo, la ventana de tiempo de trabajo de estos adhesivos dentales es mucho más amplio y ajustable.

Watch this Scientific Journal Video about Application of Light-cured Dental Adhesive Resin for Mounting Electrodes or Microdialysis Probes in Chronic Experiments at JoVE.com

Application of Light-cured Dental Adhesive Resin for Mounting Electrodes or Microdialysis Probes in Chronic Experiments

En este informe, proponemos una nueva aplicación de la luz de curado de resinas dentales base de montaje de los electrodos o sondas de microdiálisis en experimentos crónicos. Este material permite contacto directo con el cráneo.

Aplicación de curado con luz Resina Adhesivo Dental para los electrodos de montaje o sondas de microdiálisis en experimentos crónica

In chronic recording experiments, self-curing dental acrylic resins have been used as a mounting base of electrodes or microdialysis-probes. Since these ...

application-light-cured-dental-adhesive-resin-for-mounting-electrodes

Research

JoVE Journal

Biology

15.3K vistas.  RIKEN. En este informe, proponemos una nueva aplicación de la luz de curado de resinas dentales base de montaje de los electrodos o sondas de microdiálisis en experimentos crónicos. Este material permite contacto directo con el cráneo.

Video: Application of Light-cured Dental Adhesive Resin for Mounting Electrodes or Microdialysis Probes in Chronic Experiments

Preparation of Artificial Bilayers for Electrophysiology Experiments

Planar lipid bilayers, also called artificial lipid bilayers, allow you to study ion-conducting channels in a well-defined environment. These bilayers can be used for many different studies, such as the characterization of membrane-active peptides, the reconstitution of ion channels or investigations on how changes in lipid bilayer properties alter the function of bilayer-spanning channels.  Here, we show how to form a planar bilayer and how to isolate small patches from the bilayer, and in a second video  will also demonstrate a procedure for using gramicidin channels to determine changes in lipid bilayer elastic properties.   We also demonstrate the individual steps needed to prepare the bilayer chamber, the electrodes and how to test that the bilayer is suitable for single-channel measurements.

Planar lipid bilayers, also called artificial lipid bilayers, allow you to study ion-conducting channels in a well-defined environment. Here, we demonstrate the individual steps needed to prepare the bilayer chamber, the electrodes and how to test that the bilayer is suitable for single-channel measurements.

Preparación de la bicapa artificial para experimentos de electrofisiología

Planar lipid bilayers, also called artificial lipid bilayers, allow you to study ion-conducting channels in a well-defined environment. These bilayers can ...

Investigación

Biología

Fabrication of Amperometric Electrodes

Carbon fiber electrodes are crucial for the detection of catecholamine release from vesicles in single cells for amperometry measurements.  Here, we describe the techniques needed to generate low noise (&lt;0.5 pA) electrodes.  The techniques have been modified from published descriptions by previous researchers (1,2).  Electrodes are made by preparing carbon fibers and threading them individually into each capillary tube by using a vacuum with a filter to aspirate the fiber.  Next, the capillary tube with fiber is pulled by an electrode puller, creating two halves, each with a fine-pointed tip.  The electrodes are dipped in hot, liquid epoxy mixed with hardener to create an epoxy-glass seal.  Lastly, the electrodes are placed in an oven to cure the epoxy.  Careful handling of the electrodes is critical to ensure that they are made consistently and without damage.  This protocol shows how to fabricate and cut amperometric electrodes for recording from single cells.

This protocol describes how to generate carbon fiber electrodes. The electrodes are subsequently used to detect catecholamine release from vesicles with carbon fiber amperometry.

Fabricación de electrodos amperométricos

Carbon fiber electrodes are crucial for the detection of catecholamine release from vesicles in single cells for amperometry measurements.  Here, we ...

Robust 3D DNA FISH Using Directly Labeled Probes

3D DNA FISH has become a major tool for analyzing three-dimensional organization of the nucleus, and several variations of the technique have been published. In this article we describe a protocol which has been optimized for robustness, reproducibility, and ease of use. Brightly fluorescent directly labeled probes are generated by nick-translation with amino-allyldUTP followed by chemical coupling of the dye. 3D DNA FISH is performed using a freeze-thaw step for cell permeabilization and a heating step for simultaneous denaturation of probe and nuclear DNA. The protocol is applicable to a range of cell types and a variety of probes (BACs, plasmids, fosmids, or Whole Chromosome Paints) and allows for high-throughput automated imaging. With this method we routinely investigate nuclear localization of up to three chromosomal regions.

We describe a robust and versatile protocol for analyzing nuclear architecture by 3D DNA FISH using directly labeled fluorescent probes.

Robusto PESCADO ADN 3D utilizando sondas marcadas directamente

3D DNA FISH has become a major tool for  analyzing three-dimensional organization of the nucleus, and several variations  of the technique have been ...

Multilayer Mounting for Long-term Light Sheet Microscopy of Zebrafish

Light sheet microscopy is the ideal imaging technique to study zebrafish embryonic development. Due to minimal photo-toxicity and bleaching, it is particularly suited for long-term time-lapse imaging over many hours up to several days. However, an appropriate sample mounting strategy is needed that offers both confinement and normal development of the sample. Multilayer mounting, a new embedding technique using low-concentration agarose in optically clear tubes, now overcomes this limitation and unleashes the full potential of light sheet microscopy for real-time developmental biology.

The development of zebrafish can be followed over days with light sheet microscopy when embryos are embedded in optically clear polymer tubes with low-concentration agarose.

Multicapa de montaje para microscopía de luz Hoja de largo plazo del pez cebra

Light sheet microscopy is the ideal imaging technique to study zebrafish embryonic development. Due to minimal photo-toxicity and bleaching, it is ...

Isolation and Culture of Dental Epithelial Stem Cells from the Adult Mouse Incisor

Understanding the cellular and molecular mechanisms that underlie tooth regeneration and renewal has become a topic of great interest1-4, and the mouse incisor provides a model for these processes. This remarkable organ grows continuously throughout the animal&#39;s life and generates all the necessary cell types from active pools of adult stem cells housed in the labial (toward the lip) and lingual (toward the tongue) cervical loop (CL) regions. Only the dental stem cells from the labial CL give rise to ameloblasts that generate enamel, the outer covering of teeth, on the labial surface. This asymmetric enamel formation allows abrasion at the incisor tip, and progenitors and stem cells in the proximal incisor ensure that the dental tissues are constantly replenished. The ability to isolate and grow these progenitor or stem cells in vitro allows their expansion and opens doors to numerous experiments not achievable in vivo, such as high throughput testing of potential stem cell regulatory factors. Here, we describe and demonstrate a reliable and consistent method to culture cells from the labial CL of the mouse incisor.

The continuously growing mouse incisor provides a model for studying renewal of dental tissues from dental epithelial stem cells (DESCs). A robust system for consistently and reliably obtaining these cells from the incisor and expanding them in vitro is reported here.

Aislamiento y cultivo de células madre epiteliales dental del adulto Ratón Incisivo

Understanding the cellular and molecular mechanisms that underlie tooth regeneration and renewal has become a topic of great interest1-4, and the mouse ...

The Assembly and Application of 'Shear Rings': A Novel Endothelial Model for Orbital, Unidirectional and Periodic Fluid Flow and Shear Stress

Deviations from normal levels and patterns of vascular fluid shear play important roles in vascular physiology and pathophysiology by inducing adaptive as well as pathological changes in endothelial phenotype and gene expression. In particular, maladaptive effects of periodic, unidirectional flow induced shear stress can trigger a variety of effects on several vascular cell types, particularly endothelial cells. While by now endothelial cells from diverse anatomic origins have been cultured, in-depth analyses of their responses to fluid shear have been hampered by the relative complexity of shear models (e.g., parallel plate flow chamber, cone and plate flow model). While these all represent excellent approaches, such models are technically complicated and suffer from drawbacks including relatively lengthy and complex setup time, low surface areas, requirements for pumps and pressurization often requiring sealants and gaskets, creating challenges to both maintenance of sterility and an inability to run multiple experiments. However, if higher throughput models of flow and shear were available, greater progress on vascular endothelial shear responses, particularly periodic shear research at the molecular level, might be more rapidly advanced. Here, we describe the construction and use of shear rings: a novel, simple-to-assemble, and inexpensive tissue culture model with a relatively large surface area that easily allows for a high number of experimental replicates in unidirectional, periodic shear stress studies on endothelial cells.

The Assembly and Application of &#039;Shear Rings&#039;: A Novel Endothelial Model for Orbital, Unidirectional and Periodic Fluid Flow and Shear Stress

Different levels and patterns of fluid shear are known to modulate endothelial gene expression, phenotype and susceptibility to disease. We discuss the assembly and use of 'shear rings': a model that produces unidirectional, periodic shear stress patterns. Shear rings are simple to assemble, economical and can produce high cell yields.

La Asamblea y la Aplicación de &#39;Shear Anillos&#39;: A Novel endotelial Modelo de Orbital, unidireccional y periódica flujo de fluido y la tensión de cizallamiento

Deviations from normal levels and patterns of vascular fluid shear play important roles in vascular physiology and pathophysiology by inducing adaptive as ...

Application of Genetically Encoded Fluorescent Nitric Oxide (NO&#8226;) Probes, the geNOps, for Real-time Imaging of NO&#8226; Signals in Single Cells

Nitric Oxide (NO&#8226;) is a small radical, which mediates multiple important cellular functions in mammals, bacteria and plants. Despite the existence of a large number of methods for detecting NO&#8226; in vivo and in vitro, the real-time monitoring of NO&#8226; at the single-cell level is very challenging. The physiological or pathological effects of NO&#8226; are determined by the actual concentration and dwell time of this radical. Accordingly, methods that allow the single-cell detection of NO&#8226; are highly desirable. Recently, we expanded the pallet of NO&#8226; indicators by introducing single fluorescent protein-based genetically encoded nitric oxide (NO&#8226;) probes (geNOps) that directly respond to cellular NO&#8226; fluctuations and, hence, addresses this need. Here we demonstrate the usage of geNOps to assess intracellular NO&#8226; signals in response to two different chemical NO&#8226;-liberating molecules. Our results also confirm that freshly prepared 3-(2-hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-propanamine (NOC-7) has a much higher potential to evoke change in intracellular NO&#8226; levels as compared with the inorganic NO&#8226; donor sodium nitroprusside (SNP). Furthermore, dual-color live-cell imaging using the green geNOps (G-geNOp) and the chemical Ca2+ indicator fura-2 was performed to visualize the tight regulation of Ca2+-dependent NO&#8226; formation in single endothelial cells. These representative experiments demonstrate that geNOps are suitable tools to investigate the real-time generation and degradation of single-cell NO&#8226; signals in diverse experimental setups.

Application of Genetically Encoded Fluorescent Nitric Oxide (NO&amp;#8226;) Probes, the geNOps, for Real-time Imaging of NO&amp;#8226; Signals in Single Cells

This manuscript presents protocols for the application of novel genetically encoded nitric oxide (NO&#8226;) probes (geNOps) to monitor single cell NO&#8226; fluctuations in real-time using fluorescence microscopy. The Ca2+-triggered NO&#8226; formation on the level of individual endothelial cells was visualized by combining geNOps with a chemical Ca2+ sensor.

Aplicación de la codificados genéticamente fluorescente óxido nítrico (NO •) Probes, los geNOps, para imágenes en tiempo real de NO • Las señales en las células individuales

Nitric Oxide (NO&#8226;) is a small radical, which mediates multiple important cellular functions in mammals, bacteria and plants. Despite the existence ...

Inducing Apical Periodontitis in Mice

The mechanisms involved in local induced inflammation can be studied using several available animal models. One of these is the induction of apical periodontitis (AP). Apical periodontitis is a common pathology of an inflammatory nature in the periodontal tissues surrounding the tooth root. In order to better understand the nature and mechanism of this pathology it is advantageous to perform the procedure in mice. The induction of this odontogenic inflammation is achieved by drilling into the mouse tooth until the dental pulp is exposed. Next, the tooth pulp remains exposed to be contaminated by the natural oral flora over time, causing apical periodontitis. After this time period, the animal is sacrificed, and the tooth and the jaw bone can be analyzed in various ways. Typical analyses include micro-CT imaging (to evaluate bone resorption), histological staining, immunohistochemistry, and RNA expression. This protocol is useful for research in the field of oral biology to better understand this inflammatory process in an in vivo experimental setting with uniform conditions. The procedure requires a careful handling of the mice and the isolated jaw, and a visual demonstration of the technique is useful. All technical aspects of the procedures leading to induced apical periodontitis and its characterization in a mouse model are demonstrated.

Here, we present a protocol to locally induce apical periodontitis in mice. We show how to drill a hole in the mouse&#39;s tooth and expose its pulp, in order to cause local inflammation. Analysis methods to investigate the nature of this inflammation, such as micro-CT and histology, are also demonstrated.

Inducir la Periodontitis Apical en Ratones

The mechanisms involved in local induced inflammation can be studied using several available animal models. One of these is the induction of apical ...

Systematic Assessment of Mammalian Skull Specimens for Dental and Temporomandibular Joint Pathology

Museum skull specimens represent a non-invasive, informative, and readily available means to study temporomandibular joint (TMJ) lesions, dental pathology, and anatomic variations in many mammalian species. Studying the teeth and jaws of an array of species can present a challenge requiring attention to detail and understanding of a species&#39; normal anatomy. In the present article, a systematic and precise protocol for examining skull specimens is discussed that has been applied to a variety of mammals to define characteristic diseases in the oromaxillofacial region. The procedure outlined is simultaneously precise, repeatable, and adaptable to the highly differing skull and tooth shapes and anatomy across species. Specifically, specimens are examined for missing teeth, periodontal disease, endodontal disease, TMJ pathology, and anatomical variations. Results gleaned from research on museum specimens may reflect the natural history, health, and disease status of individuals and species. Furthermore, these data can inform ecological and conservation research efforts, as well as the care of captive individuals.

The present protocol describes the techniques for the systematic assessment of skull specimens to characterize anatomical and developmental variations and abnormalities of the teeth, periodontal disease, endodontal disease, and temporomandibular joint pathology.

Evaluación sistemática de muestras de cráneo de mamíferos para patología de la articulación dental y temporomandibular

Museum skull specimens represent a non-invasive, informative, and readily available means to study temporomandibular joint (TMJ) lesions, dental ...

Establecimiento del cultivo primario de células madre de la pulpa dental

Primary Culture of Dental Pulp Stem Cells

The human dental pulp represents a promising multipotent stem cell reservoir with pre-eminent regenerative competence that can be harvested from an extracted tooth. The neural crest-derived ecto-mesenchymal origin of dental pulp stem cells (DPSCs) bestows a high degree of plasticity that owes to its multifaceted benefits in tissue repair and regeneration. There are various practical ways of harvesting, maintaining, and proliferating adult stem cells being investigated for their use in regenerative medicine. In this work, we demonstrate the establishment of a primary mesenchymal stem cell culture from dental tissue by the explant culture method. The isolated cells were spindle-shaped and adhered to the plastic surface of the culture plate. The phenotypic characterization of these stem cells showed positive expression of the international society of cell therapy (ISCT)-recommended cell surface markers for MSC, such as CD90, CD73, and CD105. Further, negligible expression of hematopoietic (CD45) and endothelial markers (CD34), and less than 2% expression of HLA-DR markers, confirmed the homogeneity and purity of the DPSC cultures. We further illustrated their multipotency based on differentiation to adipogenic, osteogenic, and chondrogenic lineages. We also induced these cells to differentiate into hepatic-like and neuronal-like cells by adding corresponding stimulation media. This optimized protocol will aid in the cultivation of a highly expandable population of mesenchymal stem cells to be utilized in the laboratory or for preclinical studies. Similar protocols can be incorporated into clinical setups for practicing DPSC-based treatments.

Autor destacado: Avance de la regeneración de tejidos y el modelado de enfermedades con células madre de pulpa dental

This article provides a stepwise guide to establish a primary culture of dental pulps stem cells using the explant culture method and characterization of these cells based on ICSCRT guidelines. The cells isolated by this protocol can be considered as mesenchymal stem cells for further applications.

Cultivo primario de células madre de pulpa dental

The human dental pulp represents a promising multipotent stem cell reservoir with pre-eminent regenerative competence that can be harvested from an ...