Name: adenoviral gene therapy for diabetic keratopathy: effects on wound healing and stem cell marker expression in human organ-cultured corneas and limbal epithelial cells
Uploaded: 2016-04-07T15:00:00
Description: Watch this Scientific Journal Video about Adenoviral Gene Therapy for Diabetic Keratopathy: Effects on Wound Healing and Stem Cell Marker Expression in Human Organ-cultured Corneas and Limbal Epitheli

We gratefully acknowledge financial support by NIH/NEI R01 EY13431 (AVL), CTSI grant UL 1RR033176 (AVL), and grants from the Regenerative Medicine Institute, Cedars-Cedars Medical Center.

国立疾病研究インターチェンジ（NDRI、フィラデルフィア、PA）は承諾し、死後の健康と糖尿病の人の目と角膜を供給する。 NDRIのヒト組織収集プロトコルは、経営委員会と健康の監督の国立研究所の対象によって承認されています。本研究では、承認されたシーダーズ・サイナイ医療センター施設内倫理委員会（IRB）免除プロトコルEX-1055の下で行われています。 、博士を角膜外科医のコラボレーション。 E....

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	<li>Bikbova, G., Oshitari, T., Tawada, A., Yamamoto, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corneal+changes+in+diabetes+mellitus.">Corneal changes in diabetes mellitus.</a> Curr Diabetes Rev. 8 (4), 294-302 (2012).</li><li>Calvo-Maroto, A. M., Perez-Cambrod&#237;, R. J., Albar&#225;n-Diego, C., Pons, A., Cervi&#241;o, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+quality+of+the+diabetic+eye:+a+review.">Optical quality of the diabetic eye: a review.</a> Eye (Lond). 28 (11), 1271-1280 (2014).</li><li>Tripathy, K., Chawla, R., Sharma, Y. 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A., Yu, F. S., Castro, M. G., Ljubimov, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Normalization+of+wound+healing+and+diabetic+markers+in+organ+cultured+human+diabetic+corneas+by+adenoviral+delivery+of+c-met+gene.">Normalization of wound healing and diabetic markers in organ cultured human diabetic corneas by adenoviral delivery of c-met gene.</a> Invest Ophthalmol Vis Sci. 51 (4), 1970-1980 (2010).</li><li>Saghizadeh, M., 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=Enhanced+wound+healing,+kinase+and+stem+cell+marker+expression+in+diabetic+organ-cultured+human+corneas+upon+MMP-10+and+cathepsin+F+gene+silencing.">Enhanced wound healing, kinase and stem cell marker expression in diabetic organ-cultured human corneas upon MMP-10 and cathepsin F gene silencing.</a> Invest Ophthalmol Vis Sci. 54 (13), 8172-8180 (2013).</li><li>Saghizadeh, M., Dib, C. M., Brunken, W. J., Ljubimov, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Normalization+of+wound+healing+and+stem+cell+marker+patterns+in+organ-cultured+human+diabetic+corneas+by+gene+therapy+of+limbal+cells.">Normalization of wound healing and stem cell marker patterns in organ-cultured human diabetic corneas by gene therapy of limbal cells.</a> Exp Eye Res. 129 (Dec), 66-73 (2014).</li><li>Hatchell, D. L., 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=Damage+to+the+epithelial+basement+membrane+in+the+corneas+of+diabetic+rabbits.">Damage to the epithelial basement membrane in the corneas of diabetic rabbits.</a> Arch Ophthalmol. 101 (3), 469-471 (1983).</li><li>Chung, J. H., Kim, W. K., Lee, J. S., Pae, Y. S., Kim, H. J. <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+topical+Na-hyaluronan+on+hemidesmosome+formation+in+n-heptanol-induced+corneal+injury.">Effect of topical Na-hyaluronan on hemidesmosome formation in n-heptanol-induced corneal injury.</a> Ophthalmic Res. 30 (2), 96-100 (1998).</li><li>Saghizadeh, M., 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=Adenovirus-driven+overexpression+of+proteinases+in+organ-cultured+normal+human+corneas+leads+to+diabetic-like+changes.">Adenovirus-driven overexpression of proteinases in organ-cultured normal human corneas leads to diabetic-like changes.</a> Brain Res Bull. 81 (2-3), 262-272 (2010).</li><li>Sareen, D., 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=Differentiation+of+human+limbal-derived+induced+pluripotent+stem+cells+into+limbal-like+epithelium.">Differentiation of human limbal-derived induced pluripotent stem cells into limbal-like epithelium.</a> Stem Cells Transl Med. 3 (9), 1002-1012 (2014).</li><li>Funari, V. 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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+glucose+suppresses+epidermal+growth+factor+receptor/phosphatidylinositol+3-kinase/Akt+signaling+pathway+and+attenuates+corneal+epithelial+wound+healing.">High glucose suppresses epidermal growth factor receptor/phosphatidylinositol 3-kinase/Akt signaling pathway and attenuates corneal epithelial wound healing.</a> Diabetes. 58 (5), 1077-1085 (2009).</li><li>Xu, K., Yu, F. 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X., 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=Characterization+of+limbal+stem+cell+deficiency+by+in+vivo+laser+scanning+confocal+microscopy:+a+microstructural+approach.">Characterization of limbal stem cell deficiency by in vivo laser scanning confocal microscopy: a microstructural approach.</a> Arch Ophthalmol. 130 (4), 440-445 (2012).</li><li>Lagali, 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=In+vivo+morphology+of+the+limbal+palisades+of+Vogt+correlates+with+progressive+stem+cell+deficiency+in+aniridia-related+keratopathy.">In vivo morphology of the limbal palisades of Vogt correlates with progressive stem cell deficiency in aniridia-related keratopathy.</a> Invest Ophthalmol Vis Sci. 54 (8), 5333-5342 (2013).</li></ol>

角膜が原因遺伝子送達、ならびに有効性および副作用の評価は、簡単で、その表面の場所に遺伝子治療のための理想的な組織であると思われます。しかし、この強力なアプローチの臨床翻訳はまだによる角膜疾患や遺伝子治療のターゲット24の遺伝的原因に乏しい情報に遅いです。角膜の変化を含む糖尿病の合併症は、代謝メモリ9に変換され、自然の中で主にエピジェネティ...

糖尿病性角膜疾患は、主に退行性上皮（角膜症）と神経（神経障害）の変化をもたらします。それは多くの場合、上皮創傷治癒および角膜神経削減1-4の異常によって明らかにされています。推定60から70パーセントの糖尿病患者は1,3、様々な角膜の問題を抱えています。我々の研究は、変化したのc-Metプロトオンコジーン（肝細胞増殖因子受容体）のダウンレギュレーションを含む、ヒトの糖尿病角膜における発現およびマトリックスメタロプロテイナーゼ10（MMP-10）およびカテプシンF 5,6の上方制御で複数のマーカータンパク質を同定しました。我々はまた、大幅に文書化されたヒトの糖尿病角膜にいくつかの推定上皮幹細胞マーカーの発現が低下しています。 以前の研究では、糖尿病、遅い創傷治癒を示し、ヒト糖尿病性角膜器官培養系を用いた糖尿病改変マーカーのレベルを正常化するために、アデノウイルスに基づく遺伝子治療を開発しましたマーカーの変更、およびex vivo角膜7,8と同様の細胞マーカー発現の減少を食い止めます。この変更の持続性は、エピジェネティックな代謝メモリ9の存在によるものと思われます。この培養系は、さらに、遺伝子治療のために使用しました。この治療のためのターゲットは、糖尿病性角膜における発現低下（ のc-Met癌原遺伝子）、または増加した発現（MMP-10およびカテプシンF）のいずれかでマーカーから選択されました。 アデノウイルス（AV）療法は、全器官培養角膜または唯一の強角膜周辺輪部の区画に使用しました。この区画は、角膜上皮を更新し、積極的に4,10-15創傷治癒に関与する上皮幹細胞を保有します。ここでは、プロトコルが正常及び糖尿病のヒト角膜臓器培養、上皮創傷治癒、幹細胞に富む輪部細胞培養物の単離および特徴付け、ならびにアデノウイルス、細胞および角膜形質導入するために提供されます。私たちの結果は、将来の移植のための糖尿病性角膜にマーカー発現および創傷治癒を正規化するため、この治療法の実現可能性を示しています。彼らはまた、併用療法は、糖尿病性角膜16-18通常マーカパターンと上皮の治癒を回復するための最も効果的な方法であることを示唆しています。

<table border="0" cellpadding="0" cellspacing="0" width="1111">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:384px;">minimum essential medium</td>
			<td style="width:277px;">Thermo Fisher Scientific</td>
			<td style="width:169px;">11095-080</td>
			<td style="width:281px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Optisol-GS&#160;</td>
			<td>Bausch &#38; Lomb</td>
			<td>50006-OPT</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">ABAM antibiotic-antimycotic mixture</td>
			<td>Thermo Fisher Scientific</td>
			<td>15240062</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">calf skin collagen&#160;</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>C9791</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">agar, tissue culture grade</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>A1296</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">n-heptanol</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>72954-5ML-F</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">O.C.T. compound&#160;</td>
			<td>VWR International</td>
			<td>25608-930</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Dispase II&#160;</td>
			<td>Roche Applied Science</td>
			<td>4942078001</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">keratinocyte serum-free medium (KSFM)&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td>17005042</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">EpiLife medium with calcium</td>
			<td>Thermo Fisher Scientific</td>
			<td>MEPI500CA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">N2 medium supplement, 100x</td>
			<td>Thermo Fisher Scientific</td>
			<td>17502-048</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">B27 medium supplement, 50x</td>
			<td>Thermo Fisher Scientific</td>
			<td>17504-044</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">human keratinocyte growth supplement, 100x</td>
			<td>Thermo Fisher Scientific</td>
			<td>S-001-5</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">trypsin 0.25% - EDTA 0.02% with phenol red</td>
			<td>Thermo Fisher Scientific</td>
			<td>25200056</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">trypsin 0.25% with phenol red</td>
			<td>Thermo Fisher Scientific</td>
			<td>15050065</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">soybean trypsin inhibitor&#160;</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>T6414</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">fetal bovine serum</td>
			<td>Thermo Fisher Scientific</td>
			<td>26140079</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">insulin-transferrin-selenite supplement (ITS)</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>I3146-5ML</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">antibody to keratin 14</td>
			<td>Santa Cruz Biotechnology</td>
			<td>sc-53253</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">antibody to keratin 15</td>
			<td>Santa Cruz Biotechnology</td>
			<td>sc-47697</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">antibody to keratin 17</td>
			<td>Santa Cruz Biotechnology</td>
			<td>SC-58726</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">antibody to &#916;Np63&#945;</td>
			<td>Santa Cruz Biotechnology</td>
			<td>sc-8609</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">antibody to PAX6</td>
			<td>BioLegend</td>
			<td>PRB-278P-100</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">antibody to nidogen-1</td>
			<td>R&#38;D Systems</td>
			<td>MAB2570</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">antibody to integrin &#945;3&#946;1</td>
			<td>EMD Millipore</td>
			<td>MAB1992</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">human fibronectin</td>
			<td>BD Biosciences</td>
			<td>354008</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">human laminin</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>L4445</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">human type IV collagen</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>C6745-1ML</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">adenovirus expressing MMP-10 shRNA</td>
			<td>Capital BioSciences</td>
			<td>custom made</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">adenovirus expressing cathepsin F shRNA</td>
			<td>Capital BioSciences</td>
			<td>custom made</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">adenovirus expressing scrambled shRNA and GFP</td>
			<td>Capital BioSciences</td>
			<td>custom made</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">adenovirus expressing c-met</td>
			<td>OriGene (plasmid)</td>
			<td>SC323278</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">adenovirus expressing GFP</td>
			<td>KeraFAST</td>
			<td>FVQ002</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">sildenafil citrate, 25 mg</td>
			<td>Pfizer</td>
			<td>from pharmacy</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">epidermal growth factor&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td>PHG0311</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">poly-L-lysine</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>P4707</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">polybrene</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>107689-10G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">ViraDuctin</td>
			<td>Cell Biolabs</td>
			<td>AD-200</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">ibiBoost</td>
			<td>ibidi, Germany</td>
			<td>50301</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">phosphate buffered saline (PBS)</td>
			<td>Thermo Fisher Scientific</td>
			<td>10010049</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Corning round end spatula&#160;</td>
			<td>Dow Corning</td>
			<td>3005</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">60-mm petri dishes</td>
			<td>Thermo Fisher Scientific</td>
			<td>174888</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Nunc Lab-Tek II multiwell chamber slides&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>C6807</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">200 microliter pipet tips</td>
			<td>Bioexpress</td>
			<td>P-1233-200</td>
			<td>other suppliers available</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">inverted microscope&#160;</td>
			<td>Nikon</td>
			<td>Diaphot</td>
			<td>other suppliers/models available</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">humidified CO2 incubator&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td>370 (Steri-Cycle)</td>
			<td>other suppliers/models available</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">fluorescent microscope</td>
			<td>Olympus, Japan</td>
			<td>BX-40</td>
			<td>other suppliers/models available</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">dissecting stereo microscope</td>
			<td>Leica, Germany</td>
			<td>S4 E</td>
			<td>other suppliers/models available</td>
		</tr>
	</tbody>
</table>

adenoviral gene therapy for diabetic keratopathy: effects on wound healing and stem cell marker expression in human organ-cultured corneas and limbal epithelial cells

このプロトコルの目的は、ヒトの糖尿病角膜内の分子変化を説明し、それらが器官培養角膜にアデノウイルス遺伝子治療によって軽減することができる方法を示すことです。糖尿病性角膜疾患は、角膜神経と上皮創傷治癒の頻繁な異常を伴う糖尿病の合併症です。我々はまた、ヒトの糖尿病角膜内のいくつかの推定上皮幹細胞マーカーの有意な発現変化を報告しています。これらの変化を緩和するために、アデノウイルス遺伝子治療は、正常のc-Met癌原遺伝子発現のアップレギュレーションおよび/またはプロテイナーゼマトリックスメタロプロテイナーゼ10（MMP-10）およびカテプシンF.この治療のダウンレギュレーションを用いて実施されたが、糖尿病性角膜において創傷治癒を加速します唯一の輪部幹細胞区画を形質導入した場合であっても。最良の結果は、併用治療で得られました。正規化された幹細胞の可能性のある患者の移植のために、例もoptimizatiで提示されていますポリカチオン性増強剤を使用して幹細胞に富む培養物における遺伝子導入の上。このアプローチでは、選択された遺伝子についても、角膜上皮の創傷治癒の他のメディエーターのためだけでなく、有用であると細胞機能幹よいです。

私たちは、角膜器官培養では、糖尿病のマーカーの発現の差（ 例えば 、基底膜タンパク質とインテグリンα3β1）および正常および糖尿病角膜の間の創傷治癒が保存されていることが以前に示されています。この培養系は、C-MET、MMP-10、およびカテプシンF.、糖尿病、改変されたマーカーのレベルを正常化することを目的とした遺伝子治療を行いました<p class=...

Watch this Scientific Journal Video about Adenoviral Gene Therapy for Diabetic Keratopathy: Effects on Wound Healing and Stem Cell Marker Expression in Human Organ-cultured Corneas and Limbal Epitheli

Adenoviral Gene Therapy for Diabetic Keratopathy: Effects on Wound Healing and Stem Cell Marker Expression in Human Organ-cultured Corneas and Limbal Epithelial Cells

An example of adenoviral gene therapy in the human diabetic organ-cultured corneas is presented towards the normalization of delayed wound healing and markedly reduced epithelial stem cell marker expression in these corneas. It also describes the optimization of this process in stem cell-enriched limbal epithelial cultures.

糖尿病性角膜症のためのアデノウイルス遺伝子治療：ヒト臓器培養角膜と​​輪部上皮細胞における創傷治癒に及ぼす影響と細胞マーカーの発現を幹

The goal of this protocol is to describe molecular alterations in human diabetic corneas and demonstrate how they can be alleviated by adenoviral gene ...

Research

JoVE Journal

Developmental Biology

Generation of Murine Cardiac Pacemaker Cell Aggregates Based on ES-Cell-Programming in Combination with Myh6-Promoter-Selection

MYH6プロモーター - 選択と組み合わせたES細胞のプログラミングに基づいて、マウス心臓ペースメーカー細胞凝集体の生成

Treatment of the &#8220;sick sinus syndrome&#8221; is based on artificial pacemakers. These bear hazards such as battery failure and infections. Moreover, ...

Zebrafish Keratocyte Explants to Study Collective Cell Migration and Reepithelialization in Cutaneous Wound Healing

ゼブラフィッシュ角膜実質外植片は皮膚創傷治癒における集団細胞の移動および再上皮化を研究するために

Due to their unique motile properties, fish keratocytes dissociated from explant cultures have long been used to study the mechanisms of single cell ...

Culturing Human Pluripotent and Neural Stem Cells in an Enclosed Cell Culture System for Basic and Preclinical Research

Basicおよび前臨床研究のための密閉型細胞培養系でヒト多能性および神経幹細胞を培養します

This paper describes how to use a custom manufactured, commercially available enclosed cell culture system for basic and preclinical research. Biosafety ...

Rapid, Directed Differentiation of Retinal Pigment Epithelial Cells from Human Embryonic or Induced Pluripotent Stem Cells

人間の胚または誘導多能性幹細胞からの網膜色素上皮細胞の急速な監督の分化

We describe a robust method to direct the differentiation of pluripotent stem cells into retinal pigment epithelial cells (RPE). The purpose of providing ...

Single-cell Photoconversion in Living Intact Zebrafish

生活そのままゼブラフィッシュの単一セルの光電

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

Generation of Scaffold-free, Three-dimensional Insulin Expressing Pancreatoids from Mouse Pancreatic Progenitors In Vitro

Generation of Scaffold-free, Three-dimensional Insulin Expressing Pancreatoids from Mouse Pancreatic Progenitors In Vitro

マウス膵臓前駆細胞の in Vitroからスキャフォールド フリー、三次元のインスリンを表現する Pancreatoids の生成

The pancreas is a complex organ composed of many different cell types that work together to regulate blood glucose homeostasis and digestion. These cell ...

Efficient and Scalable Directed Differentiation of Clinically Compatible Corneal Limbal Epithelial Stem Cells from Human Pluripotent Stem Cells

効率的かつスケーラブルな臨床的に互換性のある角膜輪部上皮幹細胞ひと多能性幹細胞からの分化

Corneal limbal epithelial stem cells (LESCs) are responsible for continuously renewing the corneal epithelium, and thus maintaining corneal homeostasis ...

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

体節誘導体誘導される人間からの体外世代多能性幹細胞します。

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

Hemogenic Reprogramming of Human Fibroblasts by Enforced Expression of Transcription Factors

転写因子の強制発現によるヒト線維芽細胞のヘモジェニックリプログラミング

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

Preparation of Small RNA Libraries for Sequencing from Early Mouse Embryos

初期マウス胚からのシーケンシングのための小型RNAライブラリーの調製

MicroRNAs (miRNAs) are important for the complex regulation of cell fate decisions and developmental timing. In vivo studies of the contribution of miRNAs ...

糖尿病性角膜症のためのアデノウイルス遺伝子治療：ヒト臓器培養角膜と​​輪部上皮細胞における創傷治癒に及ぼす影響と細胞マーカーの発現を幹

糖尿病性角膜症のためのアデノウイルス遺伝子治療：ヒト臓器培養角膜と輪部上皮細胞における創傷治癒に及ぼす影響と細胞マーカーの発現を幹