Name: culture and transfection of zebrafish primary cells
Uploaded: 2018-08-17T13:00:00
Description: Watch this Scientific Journal Video about Culture and Transfection of Zebrafish Primary Cells at JoVE.com

Agradecemos a T. Fritsch, A. Lobo-Asseburg, Linde de I. y S. M. Tokarski para el excelente cuidado de los animales y soporte técnico. Agradecemos a todos los miembros del laboratorio de Köster debates intensos y provechosos. Agradecemos la financiación por la Deutsche Forschungsgemeinschaft (KO 1949/5-1) y el Estado Federal de baja Sajonia, Niedersächsisches Vorab (VWZN2889).

Todos los trabajos animales descritos aquí está de acuerdo con las regulaciones legales (EU-Directiva 2010/63). Mantenimiento y manejo de peces ha sido aprobado por las autoridades locales y por el bienestar de los animales representativo de la Universidad Tecnológica de Braunschweig, Baja Sajonia Estatal Oficina de protección al consumidor y seguridad alimentaria (LAVES, Oldenburg, Alemania; BS de TU TSchB (02.05) §4 AZ.).
1. preparación de células primarias de embriones de pez cebra...

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Aquí, presentamos dos protocolos diferentes para células en cultivo primario de 2 embriones de pez cebra de PD o de cerebro de pez cebra adulto.
La preparación de las culturas de célula primaria de pez cebra de PD 2 es relativamente fácil de realizar para cualquier persona con experiencia en técnicas de cultivo de célula básica. Sin embargo, para obtener buenos resultados reproducibles, un número suficiente de embriones como material de partida es crucial (100 es el mínimo). Durante ...

El pez cebra (Danio rerio, D. rerio) es un modelo popular vertebrado para numerosos campos de investigación básica y biomédica1. Embriones de pez cebra desarrollar rápidamente ex útero, transparente y forma bajo un microscopio, lo que proporciona excelentes condiciones para el estudio del desarrollo de vertebrados en un organismo vivo. Debido a la maleabilidad genética del pez cebra2, se han establecido muchas líneas estable reportero transgénicas con expresión de diversos marcadores fluorescentes tipo-específicas de la célula que permite la observación de poblaciones celulares específicas. La comunidad de pez cebra ofrece una amplia variedad de supuesto líneas de Gal4-conductor que llevan un transgen expresando el Kal4TA4 sintético (o el GalFF de KalTA3-equivalente) gene con el dominio de unión a DNA-Gal4 de levadura fundida a la activación transcripcional viral Dominios bajo el control de potenciadores de tipo específico de célula. Estas líneas de controlador se cruzan líneas efectoras que llevan los transgenes que consta de una secuencia activadora arriba definida (UAS) fusionada a un gen reportero. La proteína Kal4TA4 se une al elemento de UAS, activando así la expresión selectiva de tipo célula del reportero gene3,4. Este enfoque permite altamente diversos estudios combinatorios de casi todos los elementos potenciador y reportero disponibles en animales dobles transgénicos.
Sin embargo, la proyección de imagen vivo profundo con enfoque en células individuales o sus contenidos subcelulares está limitada en un embrión entero y constantemente cambiante. Para hacer frente a cuestiones biológicas celulares específicas con resolución más alta, el uso de cultivos celulares a menudo es preferible. Existen algunas líneas de células de pez cebra, pero son considerados como fuertemente seleccionados5,6,7 y su propagación es a menudo desperdiciadora de tiempo. Además, todas las líneas celulares disponibles son fibroblastos derivados, limitar los experimentos con cultivos celulares a un tipo de células. Por lo tanto, establecimos un protocolo eficiente y fácil de usar para preparar células primarias directamente en embriones de pez cebra y el cerebro del pez cebra adulto, junto con métodos para aumentar la longevidad de la cultura y ampliar la diversidad de cultivo tipos de la célula. Además, presentamos un procedimiento para transfectar las células embrionarias primarias con construcciones de la expresión de los marcadores fluorescentes organelo. Por lo tanto, morfologías celulares y estructuras subcelulares pueden ser analizadas con alta resolución espacial y temporal en tipos celulares diferentes que conservan sus características clave.

<table>
	<tbody>
		<tr>
			<td>Fish lines</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>AB (wild-type)</td>
			<td></td>
			<td></td>
			<td>established by Streisinger and colleagues, available from the Zebrafish International Resource Center (ZIRC)</td>
		</tr>
		<tr>
			<td>Tg(ptf1a:eGFP)jh1</td>
			<td></td>
			<td></td>
			<td>stable transgenic line in which the enhancer of the zebrafish gene ptf1a drives expression of the fluorescent protein EGFP (Parsons et al., 2007)</td>
		</tr>
		<tr>
			<td>Tg(XITubb:DsRed)zf148</td>
			<td></td>
			<td></td>
			<td>stable transgenic line in which the Xenopus neural-specific beta tubulin promoter drives expression of the fluorescent protein DsRed&#160; (Peri and N&#252;sslein-Volhard, 2008)</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>centrifuge</td>
			<td>Eppendorf</td>
			<td></td>
			<td>model 5804 R</td>
		</tr>
		<tr>
			<td>ChemiDoc MP imaging system</td>
			<td>BioRad</td>
			<td></td>
			<td>model XRS+, used to acquire black-and-white images of Petri dishes containing 1 da embryos</td>
		</tr>
		<tr>
			<td>confocal laser scanning microscope</td>
			<td>Leica microsystems</td>
			<td></td>
			<td>model SP8, equipped with 28 &#176;C temperature box and a 63 x objective</td>
		</tr>
		<tr>
			<td>epifluorescent microscope</td>
			<td>Leica microsystems</td>
			<td></td>
			<td>model DM5500B, equipped with 28 &#176;C temperature box and a 40 x objective</td>
		</tr>
		<tr>
			<td>Gene Pulser Xcell with capacitance extender</td>
			<td>BioRad</td>
			<td>1652661</td>
			<td>electroporation device</td>
		</tr>
		<tr>
			<td>Horizontal shaker</td>
			<td>GFL</td>
			<td></td>
			<td>model 3011</td>
		</tr>
		<tr>
			<td>incubator for cell culture (28 &#176;C)</td>
			<td>Memmert</td>
			<td></td>
			<td>model incubator I</td>
		</tr>
		<tr>
			<td>incubator for embryos (28 &#176;C)</td>
			<td>Heraeus</td>
			<td></td>
			<td>type B6120</td>
		</tr>
		<tr>
			<td>light microscope</td>
			<td>Zeiss</td>
			<td></td>
			<td>model TELAVAL 31</td>
		</tr>
		<tr>
			<td>micro pipettes</td>
			<td>Gilson</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>sterile work bench</td>
			<td>Bio Base</td>
			<td></td>
			<td>with laminar flow and UV light</td>
		</tr>
		<tr>
			<td>tweezers</td>
			<td>Dumont</td>
			<td></td>
			<td>Style 5, Inox</td>
		</tr>
		<tr>
			<td>vertical tube rotator</td>
			<td>Labinco B.V.</td>
			<td></td>
			<td>model LD-79</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Software</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Image Lab Software</td>
			<td>BioRad</td>
			<td></td>
			<td>for the ChemiDoc MP imaging system from BioRad</td>
		</tr>
		<tr>
			<td>ImageJ</td>
			<td>National Institutes of Health</td>
			<td></td>
			<td>used for counting 1 dpf embryos by applying the Count particles-tool to the respective black-and-white images; Rasband, W.S., ImageJ, U. S. National Institutes of Health, Bethesda, Maryland, USA, https://imagej.nih.gov/ij/. (1997-2016).</td>
		</tr>
		<tr>
			<td>LAS X</td>
			<td>Leica Microsystems</td>
			<td></td>
			<td>for both confocal and epifluorescent microscopes from Leica Microsystems</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Plasmids</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>pCS-DCX-tdTomato</td>
			<td>K&#246;ster Lab</td>
			<td># 1599</td>
			<td>based on the backbone pCS2+ (Rupp et al., 1994)</td>
		</tr>
		<tr>
			<td>pCS-eGFP</td>
			<td>K&#246;ster Lab</td>
			<td># 7</td>
			<td>based on the backbone pCS2+ (Rupp et al., 1994)</td>
		</tr>
		<tr>
			<td>pCS-H2B-mseCFP</td>
			<td>K&#246;ster Lab</td>
			<td># 2379</td>
			<td>based on the backbone pCS2+ (Rupp et al., 1994)</td>
		</tr>
		<tr>
			<td>pCS-mClover</td>
			<td>K&#246;ster Lab</td>
			<td># 3865</td>
			<td>based on the backbone pCS2+ (Rupp et al., 1994)</td>
		</tr>
		<tr>
			<td>pCS-MitoTag-YFP</td>
			<td>K&#246;ster Lab</td>
			<td># 2199</td>
			<td>based on the backbone pCS2+ (Rupp et al., 1994)</td>
		</tr>
		<tr>
			<td>pCS-ss-RFP-KDEL</td>
			<td>K&#246;ster Lab</td>
			<td># 4330</td>
			<td>based on the backbone pCS2+ (Rupp et al., 1994)</td>
		</tr>
		<tr>
			<td>pCS-VAMP1-mCitrine</td>
			<td>K&#246;ster Lab</td>
			<td># 2291</td>
			<td>based on the backbone pCS2+ (Rupp et al., 1994)</td>
		</tr>
		<tr>
			<td>pSK-UAS:mCherry</td>
			<td>K&#246;ster Lab</td>
			<td># 1062</td>
			<td>based on the pBluescript-backbone of Stratagene</td>
		</tr>
		<tr>
			<td>Plasmid numbers refer to the database entries of the K&#246;ster lab. Plasmids are available upon request.</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Plastic and glass ware</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>BD Falcon Cell Strainer (40 &#181;m)</td>
			<td>FALCON</td>
			<td>REF 352340</td>
			<td>distributed by BD Bioscience, used as &#8220;landing net&#8221; to dip deyolked embryos into ethanol and to transfer them quickly to fresh cell culture medium</td>
		</tr>
		<tr>
			<td>1.5 mL reaction tubes</td>
			<td>Sarstedt</td>
			<td>72690550</td>
			<td></td>
		</tr>
		<tr>
			<td>24-well plate</td>
			<td>Sarstedt</td>
			<td>83.3922</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL falconic tube</td>
			<td>Sarstedt</td>
			<td>62.547.004</td>
			<td></td>
		</tr>
		<tr>
			<td>96-well plate</td>
			<td>Sarstedt</td>
			<td>83.3924.005</td>
			<td></td>
		</tr>
		<tr>
			<td>EasyStrainer (40 &#181;m)</td>
			<td>Greiner Bio-One</td>
			<td>542 040</td>
			<td>with venting slots; used to filter cells after collagenase-mediated dissociation</td>
		</tr>
		<tr>
			<td>electroporation cuvette (0.4 cm)</td>
			<td>Kisker</td>
			<td>4905022</td>
			<td></td>
		</tr>
		<tr>
			<td>glass coverslips</td>
			<td>Heinz Herenz Medizinalbedarf GmbH</td>
			<td>1051201</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope slides</td>
			<td>Thermo Fisher Scientific (Menzel Gl&#228;ser)</td>
			<td>631-0845</td>
			<td></td>
		</tr>
		<tr>
			<td>Neubauer chamber</td>
			<td>Henneberg-Sander GmbH</td>
			<td>9020-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Pasteur pipettes (plastic; 3 mL)</td>
			<td>A. Hartenstein</td>
			<td>PP05</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dishes (plastic; diameter 10 cm)</td>
			<td>Sarstedt</td>
			<td>821473</td>
			<td>for zebrafish embryos</td>
		</tr>
		<tr>
			<td>pipette tips</td>
			<td>Sarstedt</td>
			<td>Blue (1000 &#181;l): 70762; Yellow (200 &#181;l): 70760002; White (10 &#181;L): 701116</td>
			<td></td>
		</tr>
		<tr>
			<td>sterile cell culture dishes (plastic; diameter 3 cm)</td>
			<td>TPP Techno Plastic Products AG</td>
			<td>93040</td>
			<td></td>
		</tr>
		<tr>
			<td>sterile cell culture dishes (plastic; diameter 6 cm)</td>
			<td>Sarstedt</td>
			<td>72690550</td>
			<td></td>
		</tr>
		<tr>
			<td>sterile Petri dishes (plastic; diameter 10 cm)</td>
			<td>Sarstedt</td>
			<td>83.3902</td>
			<td>for brain dissection</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Chemicals and Reagents</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>sodium chloride</td>
			<td>Roth</td>
			<td>0601.1</td>
			<td></td>
		</tr>
		<tr>
			<td>4 % paraformaldehyde in 1 x PBS</td>
			<td>Sigma-Aldrich</td>
			<td>16005</td>
			<td></td>
		</tr>
		<tr>
			<td>4&#39;,6-diamidino-2-phenylindole (DAPI)</td>
			<td>Thermo Fisher Scientific</td>
			<td>D1306</td>
			<td></td>
		</tr>
		<tr>
			<td>calcium nitrate tetrahydrate</td>
			<td>Sigma-Aldrich</td>
			<td>C1396</td>
			<td></td>
		</tr>
		<tr>
			<td>ethanol p.a. 100%</td>
			<td>Sigma-Aldrich</td>
			<td>46139</td>
			<td></td>
		</tr>
		<tr>
			<td>goat &#945;-mouse IgG (Fc specific) FITC conjugated</td>
			<td>Thermo Fisher Scientific</td>
			<td>31547</td>
			<td></td>
		</tr>
		<tr>
			<td>HEPES</td>
			<td>Roth</td>
			<td>9105.4</td>
			<td></td>
		</tr>
		<tr>
			<td>high vacuum grease</td>
			<td>DOW CORNING</td>
			<td>3826-50</td>
			<td>silicon grease used for self-made glass bottom dishes</td>
		</tr>
		<tr>
			<td>magnesium sulfate heptahydrate</td>
			<td>Merck</td>
			<td>105886</td>
			<td></td>
		</tr>
		<tr>
			<td>methylene blue</td>
			<td>Serva</td>
			<td>29198.01</td>
			<td></td>
		</tr>
		<tr>
			<td>Monoclonal Anti-Tubulin, Acetylated antibody</td>
			<td>Sigma-Aldrich</td>
			<td>T6793</td>
			<td></td>
		</tr>
		<tr>
			<td>Aqua-Poly/Mount (mounting medium)</td>
			<td>Polyscience</td>
			<td>18606</td>
			<td></td>
		</tr>
		<tr>
			<td>poly-L-lysine</td>
			<td>Biochrom</td>
			<td>L 7240</td>
			<td></td>
		</tr>
		<tr>
			<td>potasssion chloride</td>
			<td>Merck</td>
			<td>104938</td>
			<td></td>
		</tr>
		<tr>
			<td>Skim milk</td>
			<td>Roth</td>
			<td>68514-61-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Texas Red-X Phalloidin</td>
			<td>Thermo Fisher Scientific</td>
			<td>T7471</td>
			<td></td>
		</tr>
		<tr>
			<td>Tricaine</td>
			<td>Sigma-Aldrich</td>
			<td>E10521</td>
			<td>Synonym: Ethyl 3-aminobenzoate methanesulfonate</td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>BioRad</td>
			<td>1610407</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypan Blue</td>
			<td>Gibco by Life Technologies</td>
			<td>15250061</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Enzymes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>collagenase (Type 2)</td>
			<td>Thermo Fisher Scientific</td>
			<td>17101015</td>
			<td>dissolve powder in cell culture medium (8 mg/mL) and sterile-filter the solution, store aliquots at -20 &#176;C</td>
		</tr>
		<tr>
			<td>pronase (from Streptomyces griseus)</td>
			<td>Roche</td>
			<td>11459643001</td>
			<td>distributed by Sigma-Aldrich, dissolve in 30% Danieau (10 mg/mL) and store aliquots at -20 &#176;C</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Medium and solutions for cell culture</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1 x PBS (Dulbecco&#39;s Phosphate Buffered Saline)</td>
			<td>Gibco by Life Technologies</td>
			<td>14190-169</td>
			<td>distributed by Thermo Fisher Scientific</td>
		</tr>
		<tr>
			<td>CO2-independent medium</td>
			<td>Gibco by Life Technologies</td>
			<td>18045054</td>
			<td>distributed by Thermo Fisher Scientific</td>
		</tr>
		<tr>
			<td>filtrated bovine serum (FBS)</td>
			<td>PAN-Biotech</td>
			<td></td>
			<td>individual batch</td>
		</tr>
		<tr>
			<td>glutamine 100 x</td>
			<td>Gibco by Life Technologies</td>
			<td>25030081</td>
			<td>distributed by Thermo Fisher Scientific</td>
		</tr>
		<tr>
			<td>Leibovitz&#39;s L-15 medium</td>
			<td>Gibco by Life Technologies</td>
			<td>11415049</td>
			<td>distributed by Thermo Fisher Scientific</td>
		</tr>
		<tr>
			<td>PenStrep (10,000 units/mL)</td>
			<td>Gibco by Life Technologies</td>
			<td>15140148</td>
			<td>distributed by Thermo Fisher Scientific</td>
		</tr>
	</tbody>
</table>

culture and transfection of zebrafish primary cells

Embriones de pez cebra son transparentes y se desarrollan con rapidez fuera de la madre, lo que permite excelente en vivo la proyección de imagen de los procesos biológicos dinámicos en un vertebrado intacto y en desarrollo. Sin embargo, la proyección de imagen detallada de las morfologías de tipos celulares distintos y estructuras subcelulares es limitada en montajes de todo. Por lo tanto, establecimos un protocolo eficiente y fácil de usar a cultura primaria en células de embriones de pez cebra y el tejido adulto.
En Resumen, 2 embriones de pez cebra de PD son dechorionated, deyolked, esterilizados y disociar a las células con colagenasa. Después de un paso de filtración, células primarias son plateadas sobre platos de fondo de cristal y cultivadas durante varios días. Nuevas culturas, tanto como largo plazo destacan los, pueden utilizarse para estudios de proyección de imagen confocal de alta resolución. La cultura contiene diferentes tipos de células, con miocitos estriados y las neuronas siendo prominentes en la capa de poly-L-lisina. A específicamente etiqueta estructuras subcelulares por proteínas del marcador fluorescente, también establecimos un protocolo de electroporación que permite la transfección del plásmido ADN en diferentes tipos de células, incluyendo neuronas. Así, en presencia del operador estímulos definidos, comportamiento de la célula compleja y dinámica intracelular de las células primarias de pez cebra puede evaluarse con alta resolución espacial y temporal. Además, utilizando el pez cebra adulto cerebro, demostramos que la técnica de disociación descrito, así como las condiciones básicas de cultivo, también trabajan para el tejido del pez cebra adulto.

Figura 1 muestra una imagen de luz transmitida de una típica cultura derivada de embriones de tipo salvaje con miocitos estriados y racimos de células neurona-como ser más abundante. Para identificar más fácilmente a ciertos tipos de células, una línea transgénica con la expresión de una proteína fluorescente tipo-específicas de la célula puede ser utilizado (figura 1 H).
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Watch this Scientific Journal Video about Culture and Transfection of Zebrafish Primary Cells at JoVE.com

Culture and Transfection of Zebrafish Primary Cells

Presentamos un protocolo eficiente y fácil de usar para la preparación de cultivos celulares primarios de células de embriones de pez cebra para transfección celular directo imágenes así como un protocolo para preparar células primarias del cerebro del pez cebra adulto.

Cultura y transfección de células primarias de pez cebra

Zebrafish embryos are transparent and develop rapidly outside the mother, thus allowing for excellent in vivo imaging of dynamic biological processes in ...

Research

JoVE Journal

Developmental Biology

Isolation and Culture of Adult Zebrafish Brain-derived Neurospheres

Aislamiento y cultivo de neuroesferas derivada de cerebro de adultos de pez cebra

The zebrafish is a highly relevant model organism for understanding the cellular and molecular mechanisms involved in neurogenesis and brain regeneration ...

Investigación

Biología del Desarrollo

Primary Endodermal Epithelial Cell Culture from the Yolk Sac Membrane of Japanese Quail Embryos

Endodérmico primario de células epiteliales Cultura de la membrana del saco vitelino de embriones de codorniz japonesa

We established an endodermal epithelial cell culture model (EEC) for studying the function of certain enzymes and proteins in mediating nutrient ...

Isolation and Characterization of Single Cells from Zebrafish Embryos

Aislamiento y caracterización de células individuales a partir de embriones de pez cebra

The zebrafish (Danio rerio) is a powerful model organism to study vertebrate development. Though many aspects of zebrafish embryonic development have been ...

Large-Scale Production of Cardiomyocytes from Human Pluripotent Stem Cells Using a Highly Reproducible Small Molecule-Based Differentiation Protocol

La producción a gran escala de los cardiomiocitos de células madre pluripotentes utilizando un pequeño protocolo de diferenciación basada en la molécula altamente reproducible

Maximizing the benefit of human pluripotent stem cells (hPSCs) for research, disease modeling, pharmaceutical and clinical applications requires robust ...

Preparation and Culture of Myogenic Precursor Cells/Primary Myoblasts from Skeletal Muscle of Adult and Aged Humans

Preparación y cultivo de células precursoras miogénica / mioblastos primarios de músculo esquelético de los seres humanos adultos y envejecidos

Skeletal muscle homeostasis depends on muscle growth (hypertrophy), atrophy and regeneration. During ageing and in several diseases, muscle wasting ...

Single-cell Photoconversion in Living Intact Zebrafish

Sola célula Photoconversion en pez cebra intacto de la vida

Animal and plant tissue is composed of distinct populations of cells. These cells interact over time to build and maintain the tissue and can cause ...

The Isolation and Culture of Primary Epicardial Cells Derived from Human Adult and Fetal Heart Specimens

El aislamiento y cultivo de células epicárdicas principales derivan de ejemplares de corazón humano adulto y Fetal

The epicardium, an epithelial cell layer covering the myocardium, has an essential role during cardiac development, as well as in the repair response of ...

Computational Analysis of the Caenorhabditis elegans Germline to Study the Distribution of Nuclei, Proteins, and the Cytoskeleton

Computational Analysis of the Caenorhabditis elegans Germline to Study the Distribution of Nuclei, Proteins, and the Cytoskeleton

Análisis computacional de lo elegans de Caenorhabditis del Germline para estudiar la distribución de núcleos, proteínas y citoesqueleto

The Caenorhabditis elegans (C. elegans) germline is used to study several biologically important processes including stem cell development, apoptosis, and ...

Induction and Validation of Cellular Senescence in Primary Human Cells

Inducción y validación de senescencia celular en las células humanas primarias

Cellular senescence is a state of permanent cell cycle arrest activated in response to different damaging stimuli. Activation of cellular senescence is a ...

In Vitro Generation of Somite Derivatives from Human Induced Pluripotent Stem Cells

En la generación de Vitro del Somite derivados humanos inducida por células madre pluripotentes

In response to signals such as WNTs, bone morphogenetic proteins (BMPs), and sonic hedgehog (SHH) secreted from surrounding tissues, somites (SMs) give ...