Name: in vitro and in vivo models to study corneal endothelial-mesenchymal transition
Uploaded: 2016-08-20T14:00:00
Description: Watch this Scientific Journal Video about In Vitro and In Vivo Models to Study Corneal Endothelial-mesenchymal Transition at JoVE.com

We thank the staff of the Second Core Lab, Department of Medical Research, National Taiwan University Hospital for their technical support.

모든 절차는 안과 및 비전 연구에서 동물의 사용을위한 비전 및 안과 문에 연구를위한 협회 부여 본 연구에서 다음과 국립 대만 대학 병원의 기관 동물 관리 및 사용위원회에 의해 승인되었습니다. 1. 격리, 차 문화의 제조 및 소 CECs의 면역 염색 <ol><li> 로컬 도살장에서 신선한 소 눈을 획득. </li><li> 3 분 동안 10 %의 v / w 포비돈 요오드 용액에 눈 소독. 인산염 완충 식염수 (PBS) 솔루션을 씻으...

<ol>
	<li>Maurice, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+location+of+the+fluid+pump+in+the+cornea.">The location of the fluid pump in the cornea.</a> J Physiol. 221 (1), 43-54 (1972).</li><li>Joyce, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proliferative+capacity+of+the+corneal+endothelium.">Proliferative capacity of the corneal endothelium.</a> Progress in Retinal and Eye Research. 22 (3), 359-389 (2003).</li><li>Morishige, N., Sonoda, K. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bullous+keratopathy+as+a+progressive+disease:+evidence+from+clinical+and+laboratory+imaging+studies.">Bullous keratopathy as a progressive disease: evidence from clinical and laboratory imaging studies.</a> Cornea. 32, 77-83 (2013).</li><li>Bates, A. K., Cheng, H., Hiorns, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pseudophakic+bullous+keratopathy:+relationship+with+endothelial+cell+density+and+use+of+a+predictive+cell+loss+model.+A+preliminary+report.">Pseudophakic bullous keratopathy: relationship with endothelial cell density and use of a predictive cell loss model. A preliminary report.</a> Curr Eye Res. 5 (5), 363-366 (1986).</li><li>Wilson, S. E., Lloyd, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidermal+growth+factor+and+its+receptor,+basic+fibroblast+growth+factor,+transforming+growth+factor+beta-1,+and+interleukin-1+alpha+messenger+RNA+production+in+human+corneal+endothelial+cells.">Epidermal growth factor and its receptor, basic fibroblast growth factor, transforming growth factor beta-1, and interleukin-1 alpha messenger RNA production in human corneal endothelial cells.</a> Invest Ophthalmol Vis Sci. 32 (10), 2747-2756 (1991).</li><li>Chen, K. H., Harris, D. L., Joyce, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TGF-beta2+in+aqueous+humor+suppresses+S-phase+entry+in+cultured+corneal+endothelial+cells.">TGF-beta2 in aqueous humor suppresses S-phase entry in cultured corneal endothelial cells.</a> Invest Ophthalmol Vis Sci. 40 (11), 2513-2519 (1999).</li><li>Senoo, T., Obara, Y., Joyce, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EDTA:+a+promoter+of+proliferation+in+human+corneal+endothelium.">EDTA: a promoter of proliferation in human corneal endothelium.</a> Invest Ophthalmol Vis Sci. 41 (10), 2930-2935 (2000).</li><li>Yue, B. Y., Sugar, J., Gilboy, J. E., Elvart, 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=Growth+of+human+corneal+endothelial+cells+in+culture.">Growth of human corneal endothelial cells in culture.</a> Invest Ophthalmol Vis Sci. 30 (2), 248-253 (1989).</li><li>Peh, G. S., Beuerman, R. W., Colman, A., Tan, D. T., Mehta, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+corneal+endothelial+cell+expansion+for+corneal+endothelium+transplantation:+an+overview.">Human corneal endothelial cell expansion for corneal endothelium transplantation: an overview.</a> Transplantation. 91 (8), 811-819 (2011).</li><li>Zhu, C., Rawe, I., Joyce, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+protein+expression+in+human+corneal+endothelial+cells+cultured+from+young+and+older+donors.">Differential protein expression in human corneal endothelial cells cultured from young and older donors.</a> Mol Vis. 14, 1805-1814 (2008).</li><li>Ko, M. K., Kay, E. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulatory+role+of+FGF-2+on+type+I+collagen+expression+during+endothelial+mesenchymal+transformation.">Regulatory role of FGF-2 on type I collagen expression during endothelial mesenchymal transformation.</a> Invest Ophthalmol Vis Sci. 46 (12), 4495-4503 (2005).</li><li>Lee, H. T., Kay, E. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FGF-2+induced+reorganization+and+disruption+of+actin+cytoskeleton+through+PI+3-kinase,+Rho,+and+Cdc42+in+corneal+endothelial+cells.">FGF-2 induced reorganization and disruption of actin cytoskeleton through PI 3-kinase, Rho, and Cdc42 in corneal endothelial cells.</a> Mol Vis. 9, 624-634 (2003).</li><li>Pipparelli, A., 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=ROCK+Inhibitor+Enhances+Adhesion+and+Wound+Healing+of+Human+Corneal+Endothelial+Cells.">ROCK Inhibitor Enhances Adhesion and Wound Healing of Human Corneal Endothelial Cells.</a> PloS one. 8 (4), e62095 (2013).</li><li>Roy, O., Leclerc, V. B., Bourget, J. M., Theriault, M., Proulx, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+the+process+of+corneal+endothelial+morphological+change+in+vitro.">Understanding the process of corneal endothelial morphological change in vitro.</a> Invest Ophthalmol Vis Sci. 56 (2), 1228-1237 (2015).</li><li>Jakobiec, F. A., Bhat, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retrocorneal+membranes:+a+comparative+immunohistochemical+analysis+of+keratocytic,+endothelial,+and+epithelial+origins.">Retrocorneal membranes: a comparative immunohistochemical analysis of keratocytic, endothelial, and epithelial origins.</a> Am J Ophthalmol. 150 (2), 230-242 (2010).</li><li>Okumura, N., 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=Involvement+of+ZEB1+and+Snail1+in+excessive+production+of+extracellular+matrix+in+Fuchs+endothelial+corneal+dystrophy.">Involvement of ZEB1 and Snail1 in excessive production of extracellular matrix in Fuchs endothelial corneal dystrophy.</a> Lab Invest. 95 (11), 1291-1304 (2015).</li><li>Okumura, N., 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=Enhancement+on+primate+corneal+endothelial+cell+survival+in+vitro+by+a+ROCK+inhibitor.">Enhancement on primate corneal endothelial cell survival in vitro by a ROCK inhibitor.</a> Invest Ophthalmol Vis Sci. 50 (8), 3680-3687 (2009).</li><li>Zhu, Y. T., Chen, H. C., Chen, S. Y., Tseng, S. C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nuclear+p120+catenin+unlocks+mitotic+block+of+contact-inhibited+human+corneal+endothelial+monolayers+without+disrupting+adherent+junctions.">Nuclear p120 catenin unlocks mitotic block of contact-inhibited human corneal endothelial monolayers without disrupting adherent junctions.</a> J Cell Sci. 125 (15), 3636-3648 (2012).</li><li>Ho, W. 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=Inhibition+of+Matrix+Metalloproteinase+Activity+Reverses+Corneal+Endothelial-Mesenchymal+Transition.">Inhibition of Matrix Metalloproteinase Activity Reverses Corneal Endothelial-Mesenchymal Transition.</a> Am J Pathol. 185 (8), 2158-2167 (2015).</li><li>Kay, E. D., Cheung, C. C., Jester, J. V., Nimni, M. E., Smith, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Type+I+collagen+and+fibronectin+synthesis+by+retrocorneal+fibrous+membrane.">Type I collagen and fibronectin synthesis by retrocorneal fibrous membrane.</a> Invest Ophthalmol Vis Sci. 22 (2), 200-212 (1982).</li><li>Li, 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=Notch+Signal+Regulates+Corneal+Endothelial-to-Mesenchymal+Transition.">Notch Signal Regulates Corneal Endothelial-to-Mesenchymal Transition.</a> Am J Pathol. 183 (3), 786-795 (2013).</li><li>Okumura, N., 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+ROCK+Inhibitor+Eye+Drop+Accelerates+Corneal+Endothelium+Wound+Healing.">The ROCK Inhibitor Eye Drop Accelerates Corneal Endothelium Wound Healing.</a> Invest Ophthalmol Vis Sci. 54 (4), 2493-2502 (2013).</li><li>Mannermaa, E., Vellonen, K. S., Urtti, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug+transport+in+corneal+epithelium+and+blood-retina+barrier:+emerging+role+of+transporters+in+ocular+pharmacokinetics.">Drug transport in corneal epithelium and blood-retina barrier: emerging role of transporters in ocular pharmacokinetics.</a> Adv Drug Deliv Rev. 58 (11), 1136-1163 (2006).</li></ol>

CECs는 세포 증식 동안 EnMT를 받아야하는 그들의 성향 알려져있다. 치료 목적 EnMT 처리를 억제하기위한 전략을 개발하려면 EnMT기구의 완전한 이해가 필요하다. 우리는 EnMT, 체외 배양 모델 쥐의 각막 내피 세포의 cryoinjury 모델, 즉 소 CEC를 조사하기 위해이 모델을 설명했다. 우리의 결과는 두 모델의 EnMT 과정을 보여 주었다. 또한, 타틴의 EnMT 억제 효과는이 두 모델은 동일한 메커니즘을...

Corneal endothelial cells (CECs) play a vital role in maintaining corneal clarity and thus visual acuity by regulating the hydration status of the corneal stroma through active pumping1. Because of the limited proliferative potential of human CECs, the cell number decreases with age, and the repair of corneal endothelial wounds following injury is usually achieved through cell enlargement and migration, rather than cell mitosis2. When the CEC count decreases below a threshold of approximately 500 cells/mm2, the dehydration status of the corneal stroma cannot be maintained, leading to bullous keratopathy and vision impairment3,4.
The limited proliferative potential of human CECs has been attributed to several factors, including reduced expression of the epidermal growth factor and its receptor in aging cells5, antiproliferative TGF&#946;2 in the aqueous humor6, and contact inhibition2,7. Although some growth factors, such as basic fibroblast growth factor (bFGF), can increase proliferation in a cultured human corneal endothelium, the culture efficiency remains limited8,9. Furthermore, CECs may undergo a phenotypic change during ex vivo expansion, resembling epithelial-mesenchymal transition (EMT)10-13. Endothelial-mesenchymal transition (EnMT) is characterized by cell junction destabilization, apical-basal polarity loss, cytoskeletal rearrangement, alpha smooth muscle actin expression, and type I collagen secretion14. All of these characteristics may abrogate the normal function of CECs, hampering the use of ex vivo cultured CECs in tissue engineering. Moreover, EnMT has been associated with the pathogenesis of several corneal endothelial diseases, including Fuchs endothelial corneal dystrophy and retrocorneal membrane formation15,16. Therefore, understanding the mechanism of EnMT may aid in manipulating the EnMT process and facilitate the regeneration of CECs to enable competent function. 
In this study, we described a method for isolating bovine CECs from the corneal button. In the primary culture in vitro, the EnMT process, including a phenotypic change, the nuclear translocation of &#946;-catenin, and EMT regulators snail and slug, was observed. We further described a method for demonstrating EnMT in vivo by using a rat corneal endothelium cryoinjury model. Using these 2 models, we demonstrated that marimastat, a broad-spectrum matrix metalloproteinase (MMP) inhibitor, can suppress the EnMT process. The described protocols facilitate the detailed analysis of the EnMT mechanism and the development of strategies for manipulating the EnMT process for further clinical application.

<table border="0" cellpadding="0" cellspacing="0" width="1447">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:695px;">trypsin</td>
			<td style="width:269px;">ThermoFisher Scientific</td>
			<td style="width:143px;">12604-013</td>
			<td style="width:340px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dulbecco&#8217;s modified Eagle medium and Ham&#39;s F12 medium</td>
			<td>ThermoFisher Scientific</td>
			<td>11330</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">fetal bovine serum</td>
			<td>ThermoFisher Scientific</td>
			<td>26140-079</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">dimethyl sulfoxide</td>
			<td>Sigma</td>
			<td>D2650</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">human epidermal growth factor</td>
			<td>ThermoFisher Scientific</td>
			<td>PHG0311</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">insulin, transferrin, selenium&#160;</td>
			<td>ThermoFisher Scientific</td>
			<td>41400-045</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">cholera toxin</td>
			<td>Sigma</td>
			<td>C8052-1MG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">gentamicin</td>
			<td>ThermoFisher Scientific</td>
			<td>15750-060</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">amphotericin B</td>
			<td>ThermoFisher Scientific</td>
			<td>15290-026</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">paraformaldehyde</td>
			<td>Electron Microscopy Sciences</td>
			<td>111219</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Triton X-100</td>
			<td>Sigma</td>
			<td>T8787&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">bovine serum albumin</td>
			<td>Sigma</td>
			<td>A7906</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">marimastat</td>
			<td>Sigma</td>
			<td>M2699-25MG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">anti-active beta-catenin antibody</td>
			<td>Millpore</td>
			<td>05-665</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">anti-snail antibody</td>
			<td>Santa cruz</td>
			<td>sc28199</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">anti-slug antibody</td>
			<td>Santa cruz</td>
			<td>sc15391</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">goat anti-mouse IgG (H+L) secondary antibody</td>
			<td>ThermoFisher Scientific</td>
			<td>A-11001</td>
			<td>for staining of ABC of bovine CECs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">goat anti-mouse IgG (H+L) secondary antibody</td>
			<td>ThermoFisher Scientific</td>
			<td>A-11003</td>
			<td>for staining of ABC of rat corneal endothelium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">goat anti-rabbit IgG (H+L) secondary antibody</td>
			<td>ThermoFisher Scientific</td>
			<td>A-11008</td>
			<td>for staining of snail and slug of bovine CECs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">antibody diluent</td>
			<td>Genemed Biotechnologies</td>
			<td>10-0001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4&#39;,6-diamidino-2-phenylindole</td>
			<td>ThermoFisher Scientific</td>
			<td>D1306</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">mounting medium</td>
			<td>Vector Laboratories</td>
			<td>H-1000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">laser scanning confocal microscope</td>
			<td>ZEISS</td>
			<td>LSM510</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">xylazine&#160;</td>
			<td>Bayer</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">tiletamine plus zolazepam</td>
			<td>Virbac</td>
			<td>N/A</td>
			<td>veterinary drug</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">proparacaine hydrochloride ophthalmic solution</td>
			<td>Alcon</td>
			<td>N/A</td>
			<td>veterinary drug</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.1% atropine</td>
			<td>Wu-Fu Laboratories Co., Ltd</td>
			<td>N/A</td>
			<td>clinical drug&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.3% gentamicin sulfate</td>
			<td>Sinphar Group</td>
			<td>N/A</td>
			<td>clinical drug&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">basic fibroblast growth factor</td>
			<td>ThermoFisher Scientific</td>
			<td>PHG0024</td>
			<td>clinical drug&#160;</td>
		</tr>
	</tbody>
</table>

in vitro and in vivo models to study corneal endothelial-mesenchymal transition

Corneal endothelial cells (CECs) play a crucial role in maintaining corneal clarity through active pumping. A reduced CEC count may lead to corneal edema and diminished visual acuity. However, human CECs are prone to compromised proliferative potential. Furthermore, stimulation of cell growth is often complicated by gradual endothelial-mesenchymal transition (EnMT). Therefore, understanding the mechanism of EnMT is necessary for facilitating the regeneration of CECs with competent function. In this study, we prepared a primary culture of bovine CECs by peeling the CECs with Descemet's membrane from the corneal button and demonstrated that bovine CECs exhibited the EnMT process, including phenotypic change, nuclear translocation of &#946;-catenin, and EMT regulators snail and slug, in the in vitro culture. Furthermore, we used a rat corneal endothelium cryoinjury model to demonstrate the EnMT process in vivo. Collectively, the in vitro primary culture of bovine CECs and in vivo rat corneal endothelium cryoinjury models offers useful platforms for investigating the mechanism of EnMT.

소 CECs의 분리 후, 세포를 시험관 내에서 배양 하였다. (1)가 소 CECs의 위상차 이미지를 보여주고있다. 합류 셀의 육각형은 세포가 세포 분리 동안 각막의 간질 섬유 아세포에 의해 오염되지 않은 것으로 나타났다. (2)는 지정된 시점에서 ABC, 달팽이와 민달팽이에 대한 항체를 이용하여 수행 한 면역 염색을 보여주고있다.</s...

Watch this Scientific Journal Video about In Vitro and In Vivo Models to Study Corneal Endothelial-mesenchymal Transition at JoVE.com

In Vitro and In Vivo Models to Study Corneal Endothelial-mesenchymal Transition

소 각막 내피 세포의 일차 배양 각막 내피 간엽 전이의 메커니즘을 조사 하였다. 또한, 래트 각막 내피 cryoinjury 모델 생체 내 각막 내피 간엽 전환을 증명 하였다.

생체 외 및 생체 내 모델에서 각막 내피 세포 - 중간 엽 전이를 공부하기

Corneal endothelial cells (CECs) play a crucial role in maintaining corneal clarity through active pumping. A reduced CEC count may lead to corneal edema ...

Research

JoVE Journal

Developmental Biology

In Vitro and In Vivo Models to Study Corneal Endothelial-mesenchymal Transition

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

흉선의 특성 사용하여 마우스 배아에서 전구 세포를 정착<em&gt; 생체</em&gt;와<em&gt; 체외</em&gt; 분석 실험

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

연구

발생학

Stem cell-like Xenopus Embryonic Explants to Study Early Neural Developmental Features In Vitro and In Vivo

Stem cell-like Xenopus Embryonic Explants to Study Early Neural Developmental Features In Vitro and In Vivo

줄기 세포처럼<em&gt; Xenopus의</em&gt; 배아의 Explants는 초기 신경 발달의 특징을 연구하는<em&gt; 체외</em&gt; 및<em&gt; 생체</em

Understanding the genetic programs underlying neural development is an important goal of developmental and stem cell biology. In the amphibian blastula, ...

Development of an In Vitro Assay to Evaluate Contractile Function of Mesenchymal Cells that Underwent Epithelial-Mesenchymal Transition

Development of an In Vitro Assay to Evaluate Contractile Function of Mesenchymal Cells that Underwent Epithelial-Mesenchymal Transition

개발<em&gt; 체외</em&gt; 분석은 중간 엽 세포를 시행 한 상피 - 중간 엽 전환의 수축 기능을 평가하는

Fibrosis is often involved in the pathogenesis of various chronic progressive diseases such as interstitial pulmonary disease. Pathological hallmark is ...

Live Imaging of Mouse Secondary Palate Fusion

Mouse Secondary Palate Fusion의 라이브 이미징

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 ...

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains

해 부 및 Immunofluorescent 얼룩의 버섯 몸과 성인 초파리 melanogaster 두뇌 신경 세포 포토 리셉터

Nervous system development involves a sequential series of events that are coordinated by several signaling pathways and regulatory networks. Many of the ...

Corneal Tissue Engineering: An In Vitro Model of the Stromal-nerve Interactions of the Human Cornea

Corneal Tissue Engineering: An In Vitro Model of the Stromal-nerve Interactions of the Human Cornea

각 막 조직 공학:는 생체 외에서 의 모델 인간 각 막의 실질 신경 상호 작용

Tissue engineering has gained substantial recognition due to the high demand for human cornea replacements with an estimated 10 million people worldwide ...

Application of RNAi and Heat-shock-induced Transcription Factor Expression to Reprogram Germ Cells to Neurons in C. elegans

Application of RNAi and Heat-shock-induced Transcription Factor Expression to Reprogram Germ Cells to Neurons in C. elegans

C. 선 충 에서 신경 세포를 세균 세포를 재 설 정할 RNAi 및 열 충격 유도 하는 전사 인자 식의 적용

Studying the cell biological processes during converting the identities of specific cell types provides important insights into mechanism that maintain ...

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

노드 및 스캐닝 전자 현미경 검사 법, 면역 형광 산 전체를 사용 하 여 Gastrulating 마우스 배아에서 Notochordal 접시를 시각화

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

Adult Zebrafish Injury Models to Study the Effects of Prednisolone in Regenerating Bone Tissue

성인 Zebrafish 부상 모델 뼈 조직 재생에 Prednisolone의 효과 연구

Zebrafish are able to regenerate various organs, including appendages (fins) after amputation. This involves the regeneration of bone, which regrows ...

An In Vivo Method to Study Mouse Blood-Testis Barrier Integrity

An In Vivo Method to Study Mouse Blood-Testis Barrier Integrity

Vivo에서 하는 방법 마우스 혈액 고환 방 벽 완전성을 공부

Spermatogenesis is the development of spermatogonia into mature spermatozoa in the seminiferous tubules of the testis. This process is supported by ...