Name: stimulation of notch signaling in mouse osteoclast precursors
Uploaded: 2017-02-28T15:00:00
Description: Watch this Scientific Journal Video about Stimulation of Notch Signaling in Mouse Osteoclast Precursors at JoVE.com

This work was supported by the University of Pennsylvania Center for Musculoskeletal Disorders (5 P30 AR050950-09), a grant from the AO Foundation (S-16-12A), the Philadelphia VA Medical Center Translational Musculoskeletal Research Center, and an intramural orthopaedic surgery departmental research development fund. JWA is supported by the University of Pennsylvania Postdoctoral Opportunities in Research and Teaching (PENN-PORT) fellowship funded by the National Institute of General Medical Sciences Institutional Research and Career Development Award (IRACDA; 5 K12 GM081259-08).

تم إجراء كافة البحوث التي تنطوي على الحيوانات الفقارية وفقا للبروتوكولات المعتمدة من قبل جامعة ولاية بنسلفانيا رعاية الحيوان المؤسسية واللجنة الاستخدام (IACUC). 1. إعداد وسائل الإعلام الثقافة <ol style=";text-align:right;direction:rtl"><li style=";text-align:r...

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	<li>Harper, J. A., Yuan, J. S., Tan, J. B., Visan, I., Guidos, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Notch+signaling+in+development+and+disease.">Notch signaling in development and disease.</a> Clin Genet. 64 (6), 461-472 (2003).</li><li>Andersson, E. 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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PKCdelta+is+required+for+Jagged-1+induction+of+human+mesenchymal+stem+cell+osteogenic+differentiation.">PKCdelta is required for Jagged-1 induction of human mesenchymal stem cell osteogenic differentiation.</a> Stem Cells. 31 (6), 1181-1192 (2013).</li><li>Yang, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+radiation+on+the+Notch+signaling+pathway+in+osteoblasts.">Effect of radiation on the Notch signaling pathway in osteoblasts.</a> Int J Mol Med. 31 (3), 698-706 (2013).</li><li>Urs, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+soluble+Jagged1-mediated+inhibition+of+Notch+signaling+on+proliferation+and+differentiation+of+an+adipocyte+progenitor+cell+model.">Effect of soluble Jagged1-mediated inhibition of Notch signaling on proliferation and differentiation of an adipocyte progenitor cell model.</a> Adipocyte. 1 (1), 46-57 (2012).</li><li>LeComte, M. D., Shimada, I. S., Sherwin, C., Spees, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Notch1-STAT3-ETBR+signaling+axis+controls+reactive+astrocyte+proliferation+after+brain+injury.">Notch1-STAT3-ETBR signaling axis controls reactive astrocyte proliferation after brain injury.</a> Proc Natl Acad Sci U S A. 112 (28), 8726-8731 (2015).</li><li>Kostianovsky, A. M., Maier, L. M., Baecher-Allan, C., Anderson, A. C., Anderson, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Up-regulation+of+gene+related+to+anergy+in+lymphocytes+is+associated+with+Notch-mediated+human+T+cell+suppression.">Up-regulation of gene related to anergy in lymphocytes is associated with Notch-mediated human T cell suppression.</a> J Immunol. 178 (10), 6158-6163 (2007).</li><li>Teitelbaum, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Osteoclasts:+what+do+they+do+and+how+do+they+do+it?.">Osteoclasts: what do they do and how do they do it?.</a> Am J Pathol. 170 (2), 427-435 (2007).</li><li>Ma, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disruption+of+the+transcription+factor+RBP-J+results+in+osteopenia+attributable+to+attenuated+osteoclast+differentiation.">Disruption of the transcription factor RBP-J results in osteopenia attributable to attenuated osteoclast differentiation.</a> Mol Biol Rep. 40 (3), 2097-2105 (2013).</li><li>Bai, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NOTCH1+regulates+osteoclastogenesis+directly+in+osteoclast+precursors+and+indirectly+via+osteoblast+lineage+cells.">NOTCH1 regulates osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblast lineage cells.</a> J Biol Chem. 283 (10), 6509-6518 (2008).</li><li>Yamada, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+osteoclast+development+by+Notch+signaling+directed+to+osteoclast+precursors+and+through+stromal+cells.">Regulation of osteoclast development by Notch signaling directed to osteoclast precursors and through stromal cells.</a> Blood. 101 (6), 2227-2234 (2003).</li><li>Sethi, N., Dai, X., Winter, C. G., Kang, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor-derived+JAGGED1+promotes+osteolytic+bone+metastasis+of+breast+cancer+by+engaging+notch+signaling+in+bone+cells.">Tumor-derived JAGGED1 promotes osteolytic bone metastasis of breast cancer by engaging notch signaling in bone cells.</a> Cancer Cell. 19 (2), 192-205 (2011).</li><li>Sekine, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+regulation+of+osteoclastogenesis+by+Notch2/Delta-like+1+and+Notch1/Jagged1+axes.">Differential regulation of osteoclastogenesis by Notch2/Delta-like 1 and Notch1/Jagged1 axes.</a> Arthritis Res Ther. 14 (2), R45 (2012).</li><li>Ashley, J. W., Ahn, J., Hankenson, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Notch+Signaling+Promotes+Osteoclast+Maturation+and+Resorptive+Activity.">Notch Signaling Promotes Osteoclast Maturation and Resorptive Activity.</a> J Cell Biochem. 116 (11), 2598-2609 (2015).</li><li>Fukushima, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+association+of+Notch2+and+NF-kappaB+accelerates+RANKL-induced+osteoclastogenesis.">The association of Notch2 and NF-kappaB accelerates RANKL-induced osteoclastogenesis.</a> Mol Cell Biol. 28 (20), 6402-6412 (2008).</li><li>Canalis, E., Schilling, L., Yee, S. P., Lee, S. K., Zanotti, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hajdu+Cheney+Mouse+Mutants+Exhibit+Osteopenia,+Increased+Osteoclastogenesis,+and+Bone+Resorption.">Hajdu Cheney Mouse Mutants Exhibit Osteopenia, Increased Osteoclastogenesis, and Bone Resorption.</a> J Biol Chem. 291 (4), 1538-1551 (2016).</li></ol>

الخطوات الحاسمة في البروتوكول ثقافة والتمايز المختبر السلائف ناقضة العظم توفر منبرا مفيدا للتحقيق في الآليات الجزيئية من osteoclastogenesis وتحديد الأهداف العلاجية لتجديد العظام والحفاظ على كتلة العظام. عندما زراعة السلائف الماوس ?...

والثدييات الشق يشير المسار هو مثلي إلى نفس المسار في ذبابة الفاكهة، ويتكون من أربعة مستقبلات الغشاء الشق (Notch1-4) وخمسة بروابط بغشاء من خشنة (JAG1 وJAG2) ودلتا مثل (DLL1، 3، &amp; 4) أسر 1. يبدأ الشق الإشارات عندما يرتبط مستقبلات درجة على خلية المستقبلة التي يجند على خلية يحيل 2. خلال هذا التنشيط عبر والشق يجند بغشاء ينتج قوة تمتد على بغشاء الشق مستقبلات 3 و 4. القوة تمتد من يجند ملزم يؤدي الى تغييرات بتكوين في مستقبلات الشق التي تسهل انشقاق خارج الخلية من المستقبلات التي كتبها TNFalpha تحويل انزيم (TACE)، يليه حدث انشقاق داخل الخلايا بوساطة التي تحتوي على presenilin جاما secretase معقدة (γ-secretase). النشرات γ-secretase كثافة العمليات الشقنطاق racellular حوال التي translocates داخل النواة حيث يشكل مجمع تفعيل النسخي مع البرازيلي-1-سو (H) -Lag-1 (CSL)، العقل المدبر مثل (MAML)، وخلية العوامل نوع معين لدفع التعبير استهداف الجينات 5. العناصر الميكانيكية الشق يشير نتيجة التنشيط في الحاجة إلى أساليب فريدة من تفعيل الشق المسار في المختبر. يمكن بروابط الشق القابلة للذوبان ربط الشق المستقبلات، لكنه يفشل في إنتاج تمتد القوات اللازمة للافراج حوال وفي الوقت نفسه تثبيط تنافسية ملزمة بروابط الشق المرتبط الخلية. وهكذا، إضافة بروابط الشق القابلة للذوبان في ثقافة المتوسط يمكن أن تخفف الشق الطبيعي يشير 6 و 7. لحسن الحظ، يمكن بروابط الشق لحث على الإفراج حوال إذا كانت ثابتة على ركيزة صلبة مناسبة 5، 8، 9،10. بذر الخلايا على ركائز الثقافة المغلفة يجند أو تطبيق حبات المغلفة يجند إلى الخلايا على حد سواء يمكن تفعيل الشق الإشارات، والاختيار بينها يعتمد في المقام الأول على توقيت المطلوب من التحفيز الشق. لتفعيل الشق يشير فوري، مؤقت، كما هو المطلوب خلال منتصف فحص وظيفي أو التمايز، يجند الشق يمكن أن تكون ملزمة لالاغاروز الخرز، وتطبيقها على الخلايا المستزرعة، وجرفت في أي وقت. لمزيد من الإشارات الشق متواصلة من بداية الفترة الثقافة، لوحات زراعة الأنسجة يمكن أن تكون مغلفة مع يجند قبل البذر الخلية. لأغراض هذا البروتوكول، ويتم تنفيذ طرق من استخدام السلائف الماوس ناقضة العظم، ولكن الأساليب والاختلافات على الطرق الموضحة هنا تنطبق على مجموعة واسعة من أنواع الخلايا 6، 11، 12، 13، </سوب&gt; 14. الآكلة هي عضال متباينة الخلايا المكونة للدم عدد الأسطر التي هي المسؤولة عن ارتشاف النسيج العظمي، وكانوا متورطين في الاضطرابات متعددة من فقدان العظام (15). وهكذا، الدراسة في المختبر من تمايز الخلايا الآكلة من السلائف الوحيدات / بلعم-نسبهم والآليات الجزيئية التي تتحكم في وظيفتها هي ضرورية لفهم أفضل للالآكلة وتطوير علاجات جديدة لتجديد العظام. في حين اصبح من المقبول عموما أن يشير الشق تلعب دورا إيجابيا في التمايز وظيفة الخلايا الآكلة، والاختلافات في كل من التحفيز يشير الشق وناقضة العظم ثقافة السلائف والتمايز أدى إلى البداية النتائج المتناقضة 16، 17، 18، 19. دراسة أعمق للاختلافات في طرق واستخدام الوراثيةنماذج قد أوضح كثيرا من دور الإشارات الشق في osteoclastogenesis، ولكن تطبيق أساليب التحفيز الشق وثقافة موحدة يمكن منع مثل هذه الخلافات في الدراسات المستقبلية من الإشارات الشق في أنواع الخلايا الأخرى 20، 21، 22، 23. وهناك طرق متعددة لزراعة والتفريق السلائف الماوس ناقضة العظم، وكما هو الحال مع طرق مختلفة لتحفيز الشق إشارة، فإن أفضل طريقة تعتمد على مسألة تجريبية. هنا، سيتم تقديم أسلوبنا المفضل للزراعة كسور ملتصقة وغير ملتصقة من خلايا نخاع العظام مسح من الماوس طويلة. هذا الأسلوب له ميزة تتطلب في الأساس ليست معدات متخصصة وإنتاج الخلايا التي تنطبق على مجموعة متنوعة من الطرق التقليدية.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Recombinant mouse M-CSF</td>
			<td>Biolegend</td>
			<td>576402</td>
			<td>Available from multiple suppliers, test activity before experiments</td>
		</tr>
		<tr>
			<td>Recombinant mouse RANKL</td>
			<td>Shenandoah Biotechnology</td>
			<td>200-04</td>
			<td>Available from multiple suppliers, test activity before experiments</td>
		</tr>
		<tr>
			<td>Recombinant human Jagged1-Fc</td>
			<td>R&#38;D Systems</td>
			<td>1277-JG-050</td>
			<td>Available from multiple suppliers, test activity before experiments</td>
		</tr>
		<tr>
			<td>Protein G agarose beads</td>
			<td>InvivoGen</td>
			<td>gel-agg-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti-human IgG Fc</td>
			<td>Jackson ImmunoResearch</td>
			<td>109-001-008</td>
			<td></td>
		</tr>
		<tr>
			<td>Minimum Essential Medium powder</td>
			<td>Sigma-Aldrich</td>
			<td>M0894</td>
			<td></td>
		</tr>
		<tr>
			<td>Accutase cell dissociation reagent</td>
			<td>ThermoFisher</td>
			<td>A1110501</td>
			<td>Used to lift osteoclast precursors</td>
		</tr>
		<tr>
			<td>Acid Phosphatase, Leukocyte (TRAP) Kit</td>
			<td>Sigma-Aldrich</td>
			<td>387A-1KT</td>
			<td>Used to stain differentiated osteoclasts</td>
		</tr>
	</tbody>
</table>

stimulation of notch signaling in mouse osteoclast precursors

Notch signaling is a key component of multiple physiological and pathological processes. The nature of Notch signaling, however, makes in vitro investigation of its varying and sometimes contradictory roles a challenge. As a component of direct cell-cell communication with both receptors and ligands bound to the plasma membrane, Notch signaling cannot be activated in vitro by simple addition of ligands to culture media, as is possible with many other signaling pathways. Instead, Notch ligands must be presented to cells in an immobilized state. 
Variations in methods of Notch signaling activation can lead to different outcomes in cultured cells. In osteoclast precursors, in particular, differences in methods of Notch stimulation and osteoclast precursor culture and differentiation have led to disagreement over whether Notch signaling is a positive or negative regulator of osteoclast differentiation. While closer comparisons of osteoclast differentiation under different Notch stimulation conditions in vitro and genetic models have largely resolved the controversy regarding Notch signaling and osteoclasts, standardized methods of continuous and temporary stimulation of Notch signaling in cultured cells could prevent such discrepancies in the future.
This protocol describes two methods for stimulating Notch signaling specifically in cultured mouse osteoclast precursors, though these methods should be applicable to any adherent cell type with minor adjustments. The first method produces continuous stimulation of Notch signaling and involves immobilizing Notch ligand to a tissue culture surface prior to the seeding of cells. The second, which uses Notch ligand bound to agarose beads allows for temporary stimulation of Notch signaling in cells that are already adhered to a culture surface. This protocol also includes methods for detecting Notch activation in osteoclast precursors as well as representative transcriptional markers of Notch signaling activation.

والهدف من هذه الطريقة هو أن الثقافة وتنشيط الشق يشير في السلائف ناقضة العظم. عندما مثقف بشكل صحيح، السلائف ناقضة العظم يحمل التشكل ممدود في المقام الأول على شكل مغزل مع السيتوبلازم السلس (الشكل 1A). يجب توخي الحذر لتجنب تنشيط المناعة من السل?...

Watch this Scientific Journal Video about Stimulation of Notch Signaling in Mouse Osteoclast Precursors at JoVE.com

Stimulation of Notch Signaling in Mouse Osteoclast Precursors

Notch signaling is a form of cellular communication that relies upon direct contact between cells. To properly induce Notch signaling in vitro, Notch ligands must be presented to cells in an immobilized state. This protocol describes methods for in vitro stimulation of Notch signaling in mouse osteoclast precursors.

تحفيز اشارة الشق في ماوس تآكل العظام Osteoclast السلائف

Notch signaling is a key component of multiple physiological and pathological processes. The nature of Notch signaling, however, makes in vitro ...

The overall goal of this procedure is to observe the affects of notch signaling stimulation at varying points during the osteoclast differentiation process. This method can help answer key questions in the field of cell biology, such as:How does the role of notch signaling change during a cellular process? The main advantage of this technique is its marginality, which allows it to be adapted to other cells or notch ligands.While this method can provide insight into osteoclast differentiation, it can also be applied to other processes such as cell survival, proliferation, or to the differentiation of other cell types. Begin the procedure by securing a mouse in the supine position and disinfecting the skin with 70 percent ethanol. Next, make an incision along the anterior surface of one hind limb to expose the tibia and femur.Followed by an incision through the knee joint to free the distal end of the femur. Remove the femur by cutting through the end of the bone close to the pelvis and clear away any of the attached soft tissue. Cut through the ankle to remove the tibia and remove the soft tissue.Then, use forceps to hold the leg bone over a 15 milliliter conical tube and insert a 25 and five-eighths gauged needle attached to a 10 milliliter syringe into the uncut end of the bone to flush two point five milliliters of room temperature alpha MEM through the cut end of the bone into the tube. When all of the bone marrow has been removed, the bone will turn a clear, beige color. After collecting the marrow from each bone, allow any large debris to settle to the bottom of the tube and transfer the debris-free, bone marrow cell containing supernatant into a new 15 milliliter tube.Pellet the cells by centrifugation. Then lyse the red blood cells in zero point five milliliters of ACK Lysis buffer at 37 degrees Celsius for three minutes. At the end of the incubation, restore the isotonicity with 10 milliliters of PBS and collect the cells by centrifugation.Resuspend the pellet in 10 milliliters of fresh Alpha MEM for plating. In the 100 milliliter tissue culture treated dish, promouse. And incubate the cells overnight at 37 degrees Celsius in a humidified tissue-culture incubator.After enriching for the osteoclast precursors, wash the cells in five milliliters of PBS and treat them for three to five minutes with one milliliter of cell-dissociation reagent at room temperature until the osteoclast precursors can be dislodged with gentle tapping. Next, add four milliliters of Alpha MEM to the dish and transfer the cells to a 15 milliliter conical tube. After counting, dilute the precursors to a five times 10 to the fourth cells per milliliter concentration in Alpha MEM.And treat them with MCSF and RANKL, then immediately seed one milliliter of cells into each well of a 24 well tissue-treated plate. Further incubation and 37 degrees Celsius in the humidified tissue culture incubator. After two days, replace the medium with one milliliter of fresh osteoclast medium to continue the differentiation.For continuous notch signaling simulation, coat single well of a 24 well plate with 250 microliters of goat anti-human, IGG FC antibody in PBS. After one hour at room temperature, remove any unbound antibody with three washes and one milliliter of PBS. Next, add jagged1-FC fusion protein in PBS to the well for a two hour incubation at room temperature.At the end of the incubation, wash the well three times with fresh PBS as just demonstrated. After the third wash, remove the PBS and immediately plate two point six times 10 to the fourth osteoclast precursors per centimeter squared onto the coated surface. Then transfer the plate to the tissue culture incubator for continuous notch signaling for up to three days.For temporary notch signaling stimulation, first add one microgram of jagged1-FC with 40 microliters of protein G Agarose beads in one point five milliliters of PBS for two hour incubation with rotation at four degrees Celsius. At the end of the incubation, collect the beads by centrifugation followed by two washes and one point five milliliters of PBS under the same centrifuge conditions. After the second wash, re-suspend the jagged1 conjugated beads in 130 microliters per centimeter squared of culture medium and add the entire volume of beads directly onto two point six times 10 to the fourth osteoclast precursors in one well of a 24 well plate for their cold culture at 37 degrees Celsius and five percent carbon dioxide.After the appropriate experimental stimulation period, remove the beads with several washes of PBS. Notch signaling activation can then be assayed by measuring the target gene transcription. When properly cultured, osteoclast precursors exhibit a primarily elongated, spindle-shaped morphology with a smooth cytoplasm.Upon activation, the precursors spread and become flattened with foamy cytoplasm obtaining a resistance to RANK signaling and do not efficiently differentiate into mature osteoclasts. Under the correct culture and differentiation conditions, enriched osteoclast precursors will differentiate and infuse into large, TRAP-positive, multi-nuclear osteoclasts within three days of treatment. Seeding osteoclast precursors onto jagged1 FC-coated surfaces for their continuous stimulation induces the upper regulation of the expression of specific notch-target genes within 24 hours, while the expression of other notch target genes is not significantly altered by the stimulation.Temporary notch signaling stimulation, via jagged1 FC coated beads with as little as 600 nanograms of jagged1 FC protein also induces a significant increase in the expression of the specific notch target genes. While attempting this procedure, it's important to remember that a common cause of osteoclast precursor differentiation failure is by accidental activation during handling. This can be assessed by measuring specific activation markers such as TNF to Inos.After watching this video, you should have a good idea of how to differentiate osteoclast precursors and also to stimulate notch signaling, both transiently and continuously.

Research

JoVE Journal

Developmental Biology

Characterization of Thymic Settling Progenitors in the Mouse Embryo Using In Vivo and In Vitro Assays

Characterization of Thymic Settling Progenitors in the Mouse Embryo Using In Vivo and In Vitro Assays

توصيف التوتة تسوية الأسلاف في جنين الفأر عن طريق<em&gt; في فيفو</em&gt; و<em&gt; في المختبر</em&gt; فحوصات

Characterizing thymic settling progenitors is important to understand the pre-thymic stages of T cell development, essential to devise strategies for T ...

Isolation and Expansion of Adult Canine Hippocampal Neural Precursors

العزل والتوسع في تعليم الكبار الناب الحصين العصبية السلائف

The rate of neurogenesis within the adult hippocampus has been shown to vary across mammalian species. The canine hippocampus, demonstrating a structural ...

Drug Treatment and In Vivo Imaging of Osteoblast-Osteoclast Interactions in a Medaka Fish Osteoporosis Model

Drug Treatment and In Vivo Imaging of Osteoblast-Osteoclast Interactions in a Medaka Fish Osteoporosis Model

علاج المخدرات و<em&gt; في فيفو</em&gt; التصوير من خلية بناء العظم، تآكل العظام Osteoclast التفاعلات في نموذج الميداكا السمك هشاشة العظام

Bone-forming osteoblasts interact with bone-resorbing osteoclasts to coordinate the turnover of bone matrix and to control skeletal homeostasis. Medaka ...

Live Imaging of Mouse Secondary Palate Fusion

تصوير لايف من ماوس الثانوي باليت فيوجن

The fusion of the secondary palatal shelves to form the intact secondary palate is a key process in mammalian development and its disruption can lead to ...

A Novel Use of Three-dimensional High-frequency Ultrasonography for Early Pregnancy Characterization in the Mouse

استخدام الموجات فوق الصوتية عالية التردد ثلاثي الأبعاد رواية لتوصيف الحمل المبكر في الماوس

High-frequency ultrasonography (HFUS) is a common method to non-invasively monitor the real-time development of the human fetus in utero. The mouse is ...

A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation

على أساس رانكل "اوستيوكلاست الثقافة المقايسة للماوس نخاع العظام" التحقيق في دور mTORC1 في "تشكيل اوستيوكلاست"

Osteoclasts are unique bone-resorbing cells that differentiate from the monocyte/macrophage lineage of bone marrow. Dysfunction of osteoclasts may result ...

Isolation, Culture, and Differentiation of Bone Marrow Stromal Cells and Osteoclast Progenitors from Mice

العزلة، والثقافة، وتمايز الخلايا اللحمية في النخاع العظمى وفروعه اوستيوكلاست من الفئران

Bone marrow stromal cells (BMSCs) constitute a cell population routinely used as a representation of mesenchymal stem cells in vitro. They reside within ...

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands

خلية "التجميع فحوصات" لتقييم ربط الشق المورفولوجية مع ترانس-يغاندس وتثبيط عن طريق رابطة الدول المستقلة-يغاندس

Notch signaling is an evolutionarily conserved cell-cell communication system used broadly in animal development and adult maintenance. Interaction of the ...

Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes

قمع الإشارات برو تليفية يقوي التي تعالج بوساطة معامل إعادة برمجة الخلايا الليفية الجنينية الماوس في Cardiomyocytes المستحث

Trans-differentiation of one somatic cell type into another has enormous potential to model and treat human diseases. Previous studies have shown that ...

Visualizing the Node and Notochordal Plate In Gastrulating Mouse Embryos Using Scanning Electron Microscopy and Whole Mount Immunofluorescence

تصور بالعقدة ولوحة نوتوتشوردال في جاسترولاتينج الأجنة الماوس باستخدام الميكروسكوب الإلكتروني المسح وكله جبل الفلورة

The post-implantation mouse embryo undergoes major shape changes after the initiation of gastrulation and morphogenesis. A hallmark of morphogenesis is ...