Name: kinetic measurement and real time visualization of somatic reprogramming
Uploaded: 2016-07-30T13:00:00
Description: Watch this Scientific Journal Video about Kinetic Measurement and Real Time Visualization of Somatic Reprogramming at JoVE.com

De auteurs danken Chad MacArthur voor nuttige discussies.

1.Solution en Medium Voorbereiding <ol><li> Basement Membrane Matrix (gezuiverd van Engelbreth-Holm-Swarm Tumor) <ol><li> Langzaam ontdooien de basaalmembraan matrix (5 ml) op ijs bij 4 ° C overnacht. </li><li> Verdun de voorraadoplossing 1: 1 met 5 ml ijskoud, steriel DMEM / F-12 medium in een steriele, voorgekoeld 15 ml conische buis. Verdeel hoeveelheden in vooraf gekoelde, steriele 1,5 ml microcentrifuge buizen en onmiddellijk opgeslagen bij -20 ° C. </li><li> Voorafgaand aan gebruik, ontdooien ...

<ol>
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This study provides strategies for monitoring and tracking of the reprogramming process using flow cytometry and real-time imaging-based analysis. The critical steps in the protocol are initiating reprogramming, measuring reprogramming progression based on marker expression and real-time monitoring of reprogramming. Any reprogramming method of choice can be used but here we focus on Sendai based reprogramming of human fibroblasts. The advantage of this method is the ease of use and consistent high efficiency of reprogram...

-Patiënt afgeleide geïnduceerde pluripotente stamcellen (iPSCs) zijn veelbelovende instrumenten voor celtherapie en drug discovery. Zij een autologe bron van cellen voor de therapie. Bovendien, ze omvatten een zeer brede reeks genetische achtergronden, waardoor een gedetailleerde in vitro analyse van genetische ziekten dan huidige embryonale stamcel (ESC) lijnen mogelijk maken. Recente ontwikkelingen hebben geleid tot de ontwikkeling van verschillende methoden voor het genereren iPSCs, waaronder herprogrammering met Sendai-virus, episomale plasmiden of mRNA 1,2. Met name worden verschillende herprogrammering methoden in verband met verschillende niveaus van efficiëntie en veiligheid, en zullen waarschijnlijk verschillen op andere manieren hun geschiktheid van invloed zijn voor diverse toepassingen. Met de beschikbaarheid van een verscheidenheid aan technologieën herprogrammering, is het belangrijk geworden om methodologieën voor herprogrammering te ontwikkelen. De meeste bestaande werkwijzen vertrouwen op de kwalitatieve controle van de morfologie of vlekkenmet stamcel-specifieke kleurstoffen en antilichamen. Een recent ontwikkelde methode maakt gebruik van lentivirale fluorescentie reporters die gevoelig PSC-specifieke miRNAs of gedifferentieerde cel-specifieke mRNAs 3 zijn. Dergelijke controlemethoden vergemakkelijken van de selectie en de optimalisering van herprogrammering technieken voor verschillende situaties. Bijvoorbeeld is CDY1 gebruikt als een fluorescente probe te vroeg iPSCs om te screenen op modulators 4 herprogrammering. Het waarnemingsvermogen wensen herprogrammering experimenten is ook essentieel voor een beter begrip van het proces zelf. Zo is nu bekend dat bepaalde somatische celtypen zijn gemakkelijker te herprogrammeren anderen dan 5, en dat cellen doorlopen tussenfasen in herprogrammeren 6-8. Helaas, de onderliggende mechanismen herprogrammeringsproces nog niet volledig begrepen en derhalve de exacte verschillen tussen herprogrammering methoden ook nog worden defined. Zo methoden voor monitoring, evaluatie, en het vergelijken van herprogrammering gebeurtenissen blijven van cruciaal belang voor het veld stamcellen te zijn. De in dit protocol beschreven werkwijzen maken de controle en beoordeling van de herprogrammeringsproces en laten zien hoe deze technieken kunnen worden gebruikt om verschillende reeksen herprogrammering reagentia vergelijken. De eerste benadering houdt flowcytometrie geanalyseerd met behulp van combinaties van antilichamen tegen positieve en negatieve pluripotente stamcellen (PSC) markers. De tweede benadering koppels real-time beeldvorming en de meting van de totale samenloop (het percentage oppervlak bedekt door de cellen) en de samenvloeiing van marker-signalen (het percentage oppervlak bedekt door de fluorescerende signalen).

<table border="0" cellpadding="0" cellspacing="0" width="795">
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		<tr height="41">
			<td height="41" style="height:41px;width:297px;">DMEM, high glucose, GlutaMAXSupplement, pyruvate</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">10569-010</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">Fetal Bovine Serum, embryonic stem cell-qualified, US origin</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">16141-061</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">MEM Non-Essential Amino Acids Solution (100X)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">11140-050</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Trypsin-EDTA (0.05%), phenol red&#160;</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">25300-054</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">Mouse (ICR) Inactivated Embryonic Fibroblasts</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A24903</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Attachment Factor Protein (1X)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">S-006-100</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">DMEM/F-12, GlutaMAX supplement</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">10565-018</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">KnockOut Serum Replacement</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td align="right" style="width:147px;">10828010</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">2-Mercaptoethanol (55 mM)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">21985-023</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Collagenase, Type IV, powder</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">17104-019</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">TrypLE Select Enzyme (1X), no phenol red&#160;</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">12563-011</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">DPBS, no calcium, no magnesium&#160;</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">14190-144</td>
			<td></td>
		</tr>
		<tr height="61">
			<td height="61" style="height:61px;width:297px;">Geltrex LDEV-Free, hESC-Qualified, Reduced Growth Factor Basement Membrane Matrix</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A1413302</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Essential 8 Medium</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A1517001</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">FGF-Basic (AA 1-155) Recombinant Human Protein</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">PHG0264</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">UltraPure 0.5M EDTA, pH 8.0</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">15575-020</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">Bovine Albumin Fraction V (7.5% solution)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">15260-037</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">HEPES (1 M)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">15630-080</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Penicillin-Streptomycin (10,000 U/mL)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">15140-122</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">InSolution Y-27632</td>
			<td style="width:263px;">EMD Millipore</td>
			<td align="right" style="width:147px;">688001</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">CytoTune-iPS Sendai Reprogramming Kit</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A1378001</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">CytoTune-iPS 2.0 Sendai Reprogramming Kit</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A16517</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Countess II Automated Cell Counter</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">AMQAX1000</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Countess Cell Counting Chamber Slides</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">C10228</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">BJ ATCC Human Foreskin Fibroblasts, Neonatal</td>
			<td style="width:263px;">ATCC</td>
			<td style="width:147px;">CRL-2522</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">DF1 Adult Human Dermal Fibroblast</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">N/A</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">BG01V/hOG Cells Variant hESC hOct4-GFP Reporter Cells</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">R7799-105</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">IncuCyte ZOOM</td>
			<td style="width:263px;">Essen BioScience</td>
			<td style="width:147px;"></td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">SSEA-4 Antibody, Alexa Fluor 647 conjugate (MC813-70)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">SSEA421</td>
			<td></td>
		</tr>
		<tr height="61">
			<td height="61" style="height:61px;width:297px;">SSEA-4 Antibody, Alexa Fluor 488 conjugate (eBioMC-813-70 (MC-813-70))</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A14810</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">SSEA-4 Antibody (MC813-70)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">41-4000</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">TRA-1-60 Antibody (cl.A)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">&#160;41-1000</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">CD44 Rat Anti-Human/Mouse mAb (clone IM7), PE-Cy5 conjugate</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A27094</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">CD44 Alexa Fluor 488 Conjugate Kit for Live Cell Imaging</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A25528</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">CD44 Rat Anti-Human/Mouse mAb (Clone IM7)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">RM-5700 (no longer available)</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor 488 conjugate</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">&#160;A-11029</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">Goat anti-Rat IgG (H+L) Secondary Antibody, Alexa Fluor 594 conjugate</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">&#160;A-11007</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Alkaline Phosphatase Live Stain</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A14353</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">TRA-1-60 Alexa Fluor 488 Conjugate Kit for Live Cell Imaging</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A25618</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">CD24 Mouse Anti-Human mAb (clone SN3), FITC conjugate</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">MHCD2401</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">beta-2 Microglobulin Antibody, FITC conjugate (B2M-01)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A15737</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">EpCAM / CD326 Antibody, FITC conjugate (VU-1D9)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">A15755</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">CD73 / NT5E Antibody (7G2)</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">41-0200</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">VECTOR Red Alkaline Phosphatase (AP) Substrate Kit</td>
			<td style="width:263px;">Vector Laboratories</td>
			<td style="width:147px;">SK-5100</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Zeiss Axio Observer.Z1 microscope&#160;</td>
			<td style="width:263px;">Carl Zeiss</td>
			<td style="width:147px;">491912-0003-000</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">FlowJo Data Analysis Software</td>
			<td style="width:263px;">FLOJO, LLC</td>
			<td style="width:147px;">N/A</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">Attune Accoustic Focusing Cytometer, Blue/Red Laser</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td style="width:147px;">Use Attune NXT&#160;</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">S3e\ Cell Sorter (488/561 nm)</td>
			<td style="width:263px;">BIO-RAD</td>
			<td align="right" style="width:147px;">1451006</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">Falcon 12 x 75 mm Tube with Cell Strainer Cap</td>
			<td style="width:263px;">Corning</td>
			<td align="right" style="width:147px;">352235</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">Falcon 15 mL, high-clarity, dome-seal screw cap</td>
			<td style="width:263px;">Corning</td>
			<td align="right" style="width:147px;">352097</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Falcon T-75 Flask</td>
			<td style="width:263px;">Corning</td>
			<td align="right" style="width:147px;">353136</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Falcon T-175 Flask</td>
			<td style="width:263px;">Corning</td>
			<td align="right" style="width:147px;">353112</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">Falcon 6-well dish</td>
			<td style="width:263px;">Corning</td>
			<td align="right" style="width:147px;">353046</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">HERAEUS HERACELL CO2 ROLLING INCUBATOR</td>
			<td style="width:263px;">Thermo Fisher Scientific</td>
			<td align="right" style="width:147px;">51013669</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:297px;">Nonstick, RNase-free Microfuge Tubes, 1.5 mL</td>
			<td style="width:263px;"></td>
			<td style="width:147px;">AM12450</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:297px;">HulaMixer Sample Mixer</td>
			<td style="width:263px;"></td>
			<td style="width:147px;">15920D</td>
			<td></td>
		</tr>
	</tbody>
</table>

kinetic measurement and real time visualization of somatic reprogramming

Somatic reprogramming has enabled the conversion of adult cells to induced pluripotent stem cells (iPSC) from diverse genetic backgrounds and disease phenotypes. Recent advances have identified more efficient and safe methods for introduction of reprogramming factors. However, there are few tools to monitor and track the progression of reprogramming. Current methods for monitoring reprogramming rely on the qualitative inspection of morphology or staining with stem cell-specific dyes and antibodies. Tools to dissect the progression of iPSC generation can help better understand the process under different conditions from diverse cell sources. 
This study presents key approaches for kinetic measurement of reprogramming progression using flow cytometry as well as real-time monitoring via imaging. To measure the kinetics of reprogramming, flow analysis was performed at discrete time points using antibodies against positive and negative pluripotent stem cell markers. The combination of real-time visualization and flow analysis enables the quantitative study of reprogramming at different stages and provides a more accurate comparison of different systems and methods. Real-time, image-based analysis was used for the continuous monitoring of fibroblasts as they are reprogrammed in a feeder-free medium system. The kinetics of colony formation was measured based on confluence in the phase contrast or fluorescence channels after staining with live alkaline phosphatase dye or antibodies against SSEA4 or TRA-1-60. The results indicated that measurement of confluence provides semi-quantitative metrics to monitor the progression of reprogramming.

Monitoring herprogrammeren Kinetics Met behulp van flowcytometrie CD44 is een fibroblast marker terwijl SSEA4 is een PSC marker 6,10. Zoals verwacht van dit expressiepatroon flowcytometrie BJ fibroblasten toont een SSEA4 - CD44 + populatie die de creatie van kwadrant poorten maakt in combinatie met de ongekleurde monster. Tijdens de herprogrammering van DF1 fibroblasten met de Sendai v...

Watch this Scientific Journal Video about Kinetic Measurement and Real Time Visualization of Somatic Reprogramming at JoVE.com

Kinetic Measurement and Real Time Visualization of Somatic Reprogramming

Het protocol in dit onderzoek beschrijft werkwijzen voor de real-time monitoring herprogrammering progressie via de kinetische meting van positieve en negatieve pluripotente stamcel markers gebruikt flowcytometrie analyse. Het protocol bevat ook de imaging-gebaseerde beoordeling van de morfologie, en merker of reporter expressie tijdens iPSC generatie.

Kinetic Meet- en Real Time Visualisatie van Somatic herprogrammeren

Somatic reprogramming has enabled the conversion of adult cells to induced pluripotent stem cells (iPSC) from diverse genetic backgrounds and disease ...

Research

JoVE Journal

Developmental Biology

Stable and Efficient Genetic Modification of Cells in the Adult Mouse V-SVZ for the Analysis of Neural Stem Cell Autonomous and Non-autonomous Effects

Stabiele en efficiënte genetische modificatie van cellen in de Adult Mouse V-SVZ voor de Analyse van de neurale stamcel autonome en niet-autonome Effects

Relatively quiescent somatic stem cells support life-long cell renewal in most adult tissues. Neural stem cells in the adult mammalian brain are ...

Evaluation of Injury-induced Senescence and In Vivo Reprogramming in the Skeletal Muscle

Evaluation of Injury-induced Senescence and In Vivo Reprogramming in the Skeletal Muscle

Evaluatie van schade-geïnduceerde senescentie en In Vivo herprogrammering in de skeletspieren

Cellular senescence is a stress response that is characterized by a stable cellular growth arrest, which is important for many physiological and ...

Multi-Photon Time Lapse Imaging to Visualize Development in Real-time: Visualization of Migrating Neural Crest Cells in Zebrafish Embryos

Multi foton Time Lapse Imaging te visualiseren van ontwikkeling in realtime: visualisatie van migreren neurale kamcellen in Zebrafish embryo's

Congenital eye and craniofacial anomalies reflect disruptions in the neural crest, a transient population of migratory stem cells that give rise to ...

Real-time Measurement of Epithelial Barrier Permeability in Human Intestinal Organoids

Real-time meting van epitheliale barrière permeabiliteit in menselijke intestinale Organoids

Advances in 3D culture of intestinal tissues obtained through biopsy or generated from pluripotent stem cells via directed differentiation, have resulted ...

Direct Lineage Reprogramming of Adult Mouse Fibroblast to Erythroid Progenitors

Directe bloedlijn herprogrammering van volwassen muis Fibroblast te Erythroid voorlopercellen

Erythroid cell commitment and differentiation proceed through activation of a lineage-restricted transcriptional network orchestrated by a group of cell ...

Hemogenic Reprogramming of Human Fibroblasts by Enforced Expression of Transcription Factors

Hemogene herprogrammering van menselijke fibroblasten door afgedwongen expressie van transcriptiefactoren

The cellular and molecular mechanisms underlying specification of human hematopoietic stem cells (HSCs) remain elusive. Strategies to recapitulate human ...

A Layered Mounting Method for Extended Time-Lapse Confocal Microscopy of Whole Zebrafish Embryos

Een gelaagde montagemethode voor de uitgebreide time-lapse confocale microscopie van hele zebravis embryo's

Dynamics of development can be followed by confocal time-lapse microscopy of live transgenic zebrafish embryos expressing fluorescence in specific tissues ...

Real-time Bioluminescence Imaging of Notch Signaling Dynamics during Murine Neurogenesis

Real-time bioluminescentie beeldvorming van Inkeping signalering dynamiek tijdens Murine neurogenese

Notch signaling regulates the maintenance of neural stem/progenitor cells by cell-cell interactions. The components of Notch signaling exhibit dynamic ...

Real-Time Imaging of CCL5-Induced Migration of Periosteal Skeletal Stem Cells in Mice

Real-time beeldvorming van CCL5-geïnduceerde migratie van periosteale skeletstamcellen bij muizen

Periosteal skeletal stem cells (P-SSCs) are essential for lifelong bone maintenance and repair, making them an ideal focus for the development of ...

Direct Reprogramming of Mouse Fibroblasts into Melanocytes

Directe herprogrammering van muisfibroblasten in melanocyten

The loss of function of melanocytes leads to vitiligo, which seriously affects the physical and mental health of the affected individuals. Presently, ...