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

Yazarlar yararlı tartışmalar için Çad MacArthur teşekkür ederiz.

1.Solution ve Orta Hazırlık <ol><li> Bodrum Membran Matrisi (Engelbreth-Holm-Swarm Tümörü arındırılmalıdır) <ol><li> Yavaş yavaş gece boyunca 4 ° C &#39;de buz üzerinde bazal membran matrisi (5 mi) eritin. </li><li> steril, önceden soğutulmuş 15 ml konik bir tüp içinde buz soğukluğunda steril DMEM / F-12 ortamında, 5 ml 1: stok solüsyonu: 1 seyreltin. Önceden soğutulmuş, 1.5 ml steril mikro santrifüj tüplerine alikotları dağıtın ve hemen -20 ° C&#39;de saklayın. </li>...

<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...

Hasta türetilmiş uyarılmış pluripotent kök hücreler (iPSCs) hücre terapisi ve ilaç taraması için gelecek vaat eden araçlar. Onlar tedavisi için hücrelerin bir otolog kaynak sağlar. Buna ek olarak, mevcut embriyonik kök hücre (ESC) hatları sağlayacak ötesinde genetik hastalıkların detaylı in vitro analizi sağlayan genetik arka çok geniş bir dizi kapsar. Son gelişmeler, Sendai virüsü, epizomal plasmidleri ya da mRNA&#39;lar 1,2 ile yeniden programlama içeren iPSCs üretilmesi için çeşitli yöntemlerin geliştirilmesine yol açmıştır. Özellikle, farklı yeniden programlama yöntemleri verimlilik ve güvenlik çeşitli düzeylerde ilişkili ve çeşitli uygulamalar için kendi uygunluğunu etkileyen diğer yollarla farklılık muhtemel olan. yeniden programlama teknolojilerinin çeşitli kullanılabilirliği ile, yeniden programlama sürecini değerlendirmek için yöntemler geliştirmek önemli hale gelmiştir. Çoğu mevcut yöntemler morfoloji veya boyama nitel inceleme güveniyorkök hücre spesifik boyalar ve antikorlar ile. Bir son zamanlarda geliştirilen bir yöntem PSC özgü miRNA&#39;lar veya farklı hücreye özel mRNA 3 duyarlı lentiviral floresans muhabir kullanır. Böyle izleme yöntemleri seçimi ve farklı durumlar için yeniden programlama tekniklerinin optimizasyonu kolaylaştırmak. Örneğin, CDy1 yeniden programlama modülatörleri 4 taranması için erken iPSCs bir floresan sonda olarak kullanılmıştır. gözlemlemek ve farklı yeniden programlama deneyler karşılaştırmak için yeteneği de sürecin kendisi daha iyi bir anlayış kazanıyor için kritik öneme sahiptir. Örneğin, şimdi bazı somatik hücre tipleri diğerlerine 5&#39;ten yeniden programlamak kolay olduğu bilinmektedir ve hücreler 6-8 yeniden programlama sırasında ara devletler geçmesi. Ne yazık ki, yeniden programlama sürecini altında yatan mekanizmalar hala tam olarak anlaşılamamıştır ve dolayısıyla yeniden programlama yöntemleri arasında kesin farklar da d beklemektedirefined. Böylece, izleme yöntemleri, değerlendirilmesi ve yeniden programlama olayları karşılaştıran kök hücre alanı için kritik olmaya devam ediyor. Bu protokol açıklanan yöntemler izlenmesi izin ve yeniden programlama sürecinin değerlendirilmesi ve bu tekniklerin yeniden programlama reaktiflerin farklı ayarlar karşılaştırmak için nasıl kullanılabileceğini göstermektedir. İlk yaklaşım akım sitometri içeren pozitif ve negatif pluripotent kök hücre (PSC) belirteçleri karşı antikorların kombinasyonları kullanılarak analiz eder. İkinci yaklaşım çiftler gerçek zamanlı görüntüleme ve toplam izdiham (hücreler kapsadığı yüzde yüzey alanı) ve işaretleyici sinyalleri (floresan sinyalleri kapsadığı yüzde yüzey alanı) izdiham ölçümü.

<table border="0" cellpadding="0" cellspacing="0" width="795">
	<tbody>
		<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.

Akım Sitometri kullanarak yeniden programlanması Kinetiği İzleme SSEA4 PSC işaretleyici 6,10 iken CD44 bir fibroblast belirtecidir. Bu ifade modelinden beklendiği gibi, sitometri BJ fibroblast akış bir SSEA4 gösterir - boyanmamış numune ile birlikte kadran kapıları oluşturulmasını kolaylaştırır CD44 + nüfusu. SSEA4 yavaş yavaş ifade edilirken Sendai virüsler ile ...

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

Bu çalışmada sunulan protokol analizi akış sitometrisi kullanılarak, pozitif ve negatif pluripotent kök hücresi belirleyicilerinin kinetik ölçümleri ile yeniden programlama ilerlemesi gerçek zamanlı izleme için yöntemler tarif etmektedir. protokol aynı zamanda iPSC oluşturma sırasında morfolojisi ve işaretleyici veya muhabir ifade görüntüleme bazlı değerlendirmesini içerir.

Somatik yeniden programlanması Kinetik Ölçümü ve Gerçek Zamanlı Görselleştirme

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

Nöral Analizi Yetişkin Fare V-SVZ&#39;una Hücre istikrarlı ve verimli Genetik Modifikasyon Kök Hücre Özerk ve Non-özerk Etkileri

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

Yaralanma kaynaklı yaşlanma ve In Vivo kas iskelet yeniden programlama değerlendirilmesi

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

Geliştirme gerçek zamanlı olarak görselleştirmek için çoklu foton zaman atlamalı görüntüleme: nöral Crest hücrelerde geçirme Zebra balığı embriyo görselleştirme

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

Gerçek zamanlı insan bağırsak Organoids epitel bariyer geçirgenliği ölçümü

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

Lineage yetişkin fare Fibroblast Erythroid ataları için yeniden programlama doğrudan

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

Transkripsiyon Faktörlerinin Zorunlu İfadesi Ile İnsan Fibroblastlarının Hemojenik Yeniden Programlanması

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

Tüm Zebrabalığı Embriyolarının Uzun Zaman Atlamalı Konfokal Mikroskobu için Katmanlı Montaj Yöntemi

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

Murin Nörogenezi Sırasında Çentik Sinyal Dinamiğinin Gerçek Zamanlı Biyolüminesans Görüntülemesi

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

Farelerde Periosteal İskelet Kök Hücrelerinin CCL5 Kaynaklı Göçünün Gerçek Zamanlı Görüntülenmesi

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

Fare Fibroblastlarının Melanositlere Doğrudan Yeniden Programlanması

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