Name: suppression of pro-fibrotic signaling potentiates factor-mediated reprogramming of mouse embryonic fibroblasts into induced cardiomyocytes
Uploaded: 2018-06-03T14:00:00
Description: Watch this Scientific Journal Video about Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes at JoVE.com

이 연구는 Boettcher 재단의 웹 주의 생물 의학 연구 프로그램, 미국 심장 협회 과학자 개발 그랜트 (13SDG17400031), 콜로라도의 대학 학과 의학 뛰어난 초기 경력에서에서 기금에 의해 지원 되었다 학자 프로그램, 콜로라도 사단의 심장 Barlow Nyle 기부금의 대학 및 NIH R01HL133230 (캔사스)에. A.S.R 섬유 증 연구 및 번역 (CFReT)에 대 한 NIH/NCATS 콜로라도 CTSA 보조금 번호 TL1TR001081와 콜로라도 대학 컨소시엄에서 미리 박사 친목에 의해 지원 되었다. 이 연구는 암 센터 지원 그랜트 (P30CA046934), 피부 질환 연구 코어 그랜트 (P30AR057212), 그리고 콜로라도의 대학 Anschutz 의료 캠퍼스에서 Flow Cytometry 코어에 의해 또한 지원 되었다.

동물을 요구 하는 모든 실험 기관 동물 관리 및 사용 위원회 UC 덴버 Anschutz 의료 캠퍼스에 의해 승인 되었다.
1입니다. MEFs의 격리
<ol><li>E13에서 임신 마우스 C57BL/6를 구입. 밤새 배.</li>
	<li>승인 된 IACUC 프로토콜에 따라 어머니를 안락사 (예: ~1.3 L/분 CO2 동물 죽은 나타날 때까지 다음 자 궁 경관 탈 구)</li>
	<li>70% 에탄올으로 어머니를 살포 하 고 복 부 구멍을 엽니다. ?...

<ol>
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현재 연구 직접 프로 거리 신호 통로의 억제와 함께 요소를 프로그래밍 하는 GHMT2m의 배달을 통해 기능 iCMs에 섬유 아 세포를 재 설 정할 고효율 전략을 설명 합니다. Cytometry, immunofluorescent 이미징, 이미징, 및 셀 카운트, 우리는이 프로토콜에 대부분 셀의 표시 치고 칼슘을 사용 하 성공적인 프로그래밍 받 다 고 CM 혈통 운명 채택. 우리는 반대로 거리의 추가 화합물 TGF-β 유형 포함 나 수용 체 억제?...

허 혈 성 심장 질환은 미국1에서 죽음의 주요 원인. 약 800000 미국인 경험을 첫 번째 또는 재발 성 심근 경색 (MI) 당 년1. 미, 다음 cardiomyocytes (CMs) 및 심장 섬유 증, 활성화 된 심장 섬유 아 세포에서의 죽음 심장 기능2,3손상. 미 다음 심장 마비의 진행 성인 CMs4,5의 가난한 재생 용량 때문 돌이킬 아니다. 현재 임상 치료 질병의 진행을 느리게 하 고 미래의 심장 이벤트6,7,,89의 위험을 감소, 아무 치료 반대로 질병의 진행 하는 무 능력 때문 CMs 후 경색10를 다시 생성 합니다. 새로운 세포 치료 치료 환자 닉 실망 하 떠오르고 있다, 지금까지 미 다음 마음에 줄기 세포를 제공 하는 임상 시험 결정적이 아닌 재생 잠재적인11,12, 나타났습니다. 13 , 14 , 15 , 16 , 17 , 18.
먼저 다카하시 & 야 마나카, 시연 4 녹음 방송 요인의 overexpression에 의해 섬유 아 세포에서 파생 하는 인간 유도 만능 줄기 세포 (hiPSCs)의 생성 세포 치료19에 새로운 돌파구를 문을 열었습니다. 이 세포는 모든 3 개의 세균 층19로 분화 할 수 있다 그리고 CMs의 많은 수를 생성 하기 위한 몇 가지 매우 효율적인 방법을 이전 표시 되었습니다20,21. HiPSC 파생 된 CMs (엉덩이-CMs) cardiomyogenesis를 공부 하는 강력한 플랫폼을 제공 하 고 상해를 따르는 마음을 고치기를 위한 중요 한 의미를 가질 수 있습니다. 그러나, 현재 기형종 형성22의 문제로 인해 변환 장애물을 직면 하는 엉덩이 CMs 그리고 그들의 미 성숙한 성격 프로 arrhythmogenic23을 수 있습니다. HiPSCs로 섬유 아 세포를 프로그래밍 직접 다른 세포 유형으로 섬유 아 세포를 프로그래밍에 관심을 촉발 시켰다. 것 외. 시연 심장 혈통, 직접 프로그래밍에 섬유 아 세포 결과에 GATA4, Mef2c, 및 Tbx5 (그리니치 표준시)의 그 overexpression 이기는 하지만 낮은 효율24. 프로그래밍 효율 Hand2 (GHMT)25의 추가 함께 개선 되었습니다. 이러한 초기 연구 이후 많은 출판물을26,27,,2829, 요인 추가 전사와 재활 요소 칵테일 변경 증명 하고있다 chromatin 한정자30,31, microRNAs32,33또는34 향상을 리드 하는 작은 분자 프로그래밍 효율 및 성숙 유도 cardiomyocyte 같은 셀 (iCMs ).
여기 우리는 높은 효율 iCMs 마우스 미 발달 섬유 아 세포 (MEFs)에서 생성 하는 상세한 프로토콜을 제공 합니다. 우리 이전 보여주었다 GHMT 칵테일 1 미르와 미르-133 (GHMT2m)의 추가 함께 크게 개선 하 고 더욱 향상 프로-거리 경로 변형 성장 인자 (TGF-β) β 신호를 포함 하 여 신호 또는 관련 단백질 키 니 아 제 (바위) 신호 통로 저해35. 이 프로토콜을 사용 하 여, 우리 보여 심장 분 T (cTnT)를 표현 하는 세포의 약 60%, 약 50% α-actinin, 표현 및 높은 구타 셀 수 빠르면 하루 11 프로그래밍 요소와 치료의 변환에 따라 관찰 될 수 있다 TGF-β와 내가 입력 수용 체 억제제 A-83-01. 또한, 이러한 iCMs 갭 정션 단백질 connexin 43 등을 표현 하 고 자연 스러운 수축 및 칼슘 과도. 이전 연구에 비해 효율을 프로그래밍 향상을 표시이 심장 후 경색에 남아 있는 생 세포 인구에서 CMs를 재생성을 보여줍니다.

<table border="0" cellpadding="0" cellspacing="0" width="1326">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">C57BL/6 Mice</td>
			<td style="width:197px;">Charles River&#39;s Laboratory</td>
			<td style="width:111px;">027</td>
			<td style="width:667px;">For MEF isolation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Platinum E (PE) Cells</td>
			<td>Cell Biolabs, INC</td>
			<td>RV-101</td>
			<td>For retrovirus production</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DMEM High Glucose</td>
			<td>Gibco</td>
			<td>SH30022.FS</td>
			<td>Component of iCM, PE, and Growth media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Medium 199</td>
			<td>Life Technologies</td>
			<td>11150-059</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fetal Bovine Serum</td>
			<td>Gemini</td>
			<td>100106</td>
			<td>Component of iCM, PE, and Growth media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Donor Horse Serum</td>
			<td>Gemini</td>
			<td>100508 500</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">MEM Essential Amino Acids, 50X</td>
			<td>Life Technologies</td>
			<td>11130051</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Sodium Pyruvate Solution, 100X</td>
			<td>Life Technologies</td>
			<td>11360070</td>
			<td>Component of iCM media and for calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">MEM Non-Essential Amino Acids, 100X</td>
			<td>Life Technologies</td>
			<td>11140050</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">MEM Vitamin Solution, 100X</td>
			<td>Life Technologies</td>
			<td>11120-052</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Insulin-Transferrin-Selenium</td>
			<td>Gibco</td>
			<td>41400045</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">B27</td>
			<td>Gibco</td>
			<td>17504-044</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Penicilin-Streptomycin</td>
			<td>Gibco</td>
			<td>15140-122</td>
			<td>Component of iCM, PE, and Growth media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">GlutaMAX (L-Glutamine Supplement)</td>
			<td>Gibco</td>
			<td>35050-061</td>
			<td>Component of iCM, PE, and Growth media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Blasticidin-HCl</td>
			<td>Life Technologies</td>
			<td>A11139-03</td>
			<td>Component of PE media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Puromycin dihydrochloride</td>
			<td>Life Technologies</td>
			<td>A11138-03</td>
			<td>Component of PE media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">0.25% Trypsin/EDTA</td>
			<td>Gibco</td>
			<td>25200-056</td>
			<td>For detaching cells from culture dishes</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">A-83-01</td>
			<td>R&#38;D Systems - Tocris</td>
			<td>2939/10</td>
			<td>Treat cells to inhibit TGF-&#946; signaling - promotes high efficiecy reprogramming. Use at 0.5 &#181;M</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DMSO</td>
			<td>Thermo Scientific</td>
			<td>85190</td>
			<td>For dilution and storage of A-83-01 and component of Freeze Medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">SureCoat</td>
			<td>Cellutron</td>
			<td>SC-9035</td>
			<td>For coating dishes to plate MEFs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FuGENE 6 Transfection Reagent</td>
			<td>Promega</td>
			<td>E2692</td>
			<td>Transfection Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Opti-MEM Reduced Serum Media</td>
			<td>Gibco</td>
			<td>11058-021</td>
			<td>Transfection Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X Myc-GATA4</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X Myc-Hand2</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X Myc-Mef2c</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X Myc-Tbx5</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X miR-1</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X miR-133</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X GFP</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Polybrene (Hexadimethrine bromide)</td>
			<td>Sigma</td>
			<td>H9268-5G</td>
			<td>For viral induction. Use at a concentration of 6 &#181;g/mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Vacuum Filter + bottles (0.22 &#181;m pores)</td>
			<td>Nalgene</td>
			<td>&#160;569-0020&#160;</td>
			<td>For filtering media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Syringes</td>
			<td>Bd Vacutainer Labware&#160;</td>
			<td>309654</td>
			<td>For viral filtration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.45 &#181;m Filters</td>
			<td>Celltreat</td>
			<td>229749</td>
			<td>For viral filtration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">70 &#181;m cell strainers</td>
			<td>Falcon&#160;</td>
			<td>352350</td>
			<td>For MEF isolation and Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Cytofix/Cytoperm Solution</td>
			<td>BD</td>
			<td>554722</td>
			<td>For fixation and permeabilization of cells for flow cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">perm/wash buffer&#160;</td>
			<td>BD</td>
			<td>554723</td>
			<td>For washing cells for flow cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DPBS 1X</td>
			<td>Gibco</td>
			<td>14190-250</td>
			<td>For washing cells</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bovine Serum Albumin</td>
			<td>VWR</td>
			<td>0332-100g</td>
			<td>For flow cytometry and calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Goat Serum</td>
			<td>Sigma&#160;</td>
			<td>G9023</td>
			<td>For blocking cells for Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Donkey Serum</td>
			<td>Sigma</td>
			<td>D9663-10mg</td>
			<td>For blocking cells for Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mouse Troponin T</td>
			<td>Thermo Scientific</td>
			<td>ms-295-p</td>
			<td>1:400 IF, 1:200 Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mouse &#945;-actinin</td>
			<td>Sigma</td>
			<td>A7811L</td>
			<td>1:400 IF, 1:200 Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rabbit Connexin 43</td>
			<td>Sigma&#160;</td>
			<td>C6219</td>
			<td>1:400 IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rabbit Troponin I</td>
			<td>PhosphoSolutions</td>
			<td>2010-TNI</td>
			<td>1:400 IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hoechst</td>
			<td>Life Technologies</td>
			<td>62249</td>
			<td>1:10000 IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Alexa 488, rabbit</td>
			<td>Life Technologies</td>
			<td>A-11034</td>
			<td>1:800 IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Alexa 555, mouse</td>
			<td>Life Technologies</td>
			<td>A-21422</td>
			<td>1:800 IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Alexa 647, mouse</td>
			<td>Life Technologies</td>
			<td>A-31571</td>
			<td>1:200 Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">27-color ZE5 Flow Cytometer&#160;</td>
			<td>Bio-RAD</td>
			<td></td>
			<td>For FACS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Paraformaldehyde</td>
			<td>sigma</td>
			<td>P6148-500mg</td>
			<td>For fixing cells for IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Triton X-100</td>
			<td>Promega</td>
			<td>H5142</td>
			<td>For permeabilization of cells for IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">EVOS&#8482; FL Color Imaging System</td>
			<td>Thermo Scientific</td>
			<td>AMEFC4300</td>
			<td>For IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NaCl</td>
			<td>RPI</td>
			<td>S23020-5000</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">KCl</td>
			<td>VWR</td>
			<td>395</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">CaCl2</td>
			<td>Fisher</td>
			<td>C614-500</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">MgCl2</td>
			<td>VWR</td>
			<td>97061-352</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">glucose</td>
			<td>sigma</td>
			<td>G7528-250g</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">HEPES</td>
			<td>sigma</td>
			<td>H4034-500g</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fura-2 AM</td>
			<td>Life Technologies</td>
			<td>F1221</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fluronic F-127</td>
			<td>Sigma</td>
			<td>P2443-250g</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nifedipine</td>
			<td>Sigma</td>
			<td>N7634-1G</td>
			<td>For disruption calcium transients in iCMs - use at 10 &#181;M</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoproterenol</td>
			<td>sigma</td>
			<td>I6504-1g</td>
			<td>For increasing number of calcium transients in iCMs - use at 1-2 &#181;M</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Marianas Spinning Disk Confocal microscope</td>
			<td>3i</td>
			<td></td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ethanol</td>
			<td>Decon Laboratories</td>
			<td>2801</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">bleach</td>
			<td>Clorox</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">50 mL conical tubes</td>
			<td>GREINER BIO-ONE</td>
			<td>227261</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">15 mL conical tubes</td>
			<td>GREINER BIO-ONE</td>
			<td>188271</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">15 cm cell culture dishes</td>
			<td>Falcon</td>
			<td>353025</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10 cm cell culture dishes</td>
			<td>Falcon</td>
			<td>353003</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">60 mm cell culture dishes</td>
			<td>GREINER BIO-ONE</td>
			<td>628160</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">6 well cell culture plates</td>
			<td>GREINER BIO-ONE</td>
			<td>657160&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">12 well cell culture plates</td>
			<td>GREINER BIO-ONE</td>
			<td>665180&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">24 well cell culture plates</td>
			<td>GREINER BIO-ONE</td>
			<td>662160&#160;</td>
			<td></td>
		</tr>
	</tbody>
</table>

suppression of pro-fibrotic signaling potentiates factor-mediated reprogramming of mouse embryonic fibroblasts into induced cardiomyocytes

다른 하나의 체세포 형식의 트랜스-차별화 모델링 하 고 인간의 질병 치료 엄청난 잠재력이 있다. 이전 연구 결과 나타났습니다 그 마우스 배아, 피부, 및 심장 섬유 아 세포 기능 유도 cardiomyocyte 같은 세포 (iCMs)로 재설정 될 수 있습니다 cardiogenic 녹음 방송 요인 Mef2c, GATA4, Hand2 등의 overexpression 통해 고 Tbx5 둘 다 생체 외에서 그리고 에 비보. 그러나, 이러한 이전 연구는 상대적으로 낮은 효율을 나타났습니다. 부상 다음 심장 기능을 복원 하기 위해 심장 프로그래밍 제어 메커니즘 효율과 성숙 iCMs의 향상을 해명 한다.
우리는 이전 프로 거리 신호를 크게 저해 재활 효율을 증가 입증. 여기, 우리는 최대 60%의 재활 효율을 달성 하는 방법 선발. 또한, cytometry, immunofluorescent 이미징 및 칼슘 이미징 재활 효율과 재설정된 섬유 아 세포의 성숙 척도를 포함 하는 여러 가지 방법을 설명 합니다. 여기 상세한 프로토콜을 사용 하 여, 기계 연구 수 이루어져야 심장 프로그래밍의 긍정적이 고 부정적인 레 귤 레이 터를 결정. 이러한 연구는 재활 효율성과 성숙, 인간의 심장 질환 치료 소설 세포 치료로 이어질 수 있는 홍보 하는 대상으로 수 신호 경로 식별할 수 있습니다.

그림 1B에서 위에 설명 된 재활 전략을 사용 하 여, 약 심장 분 T를 표현 하는 세포의 70%와 심장 α-actinin, cytometry 하루 9에 의해 계량을 표현 하는 세포의 약 55 %iCMs 생성 GHMT2m의 변환에 따라 (그림 2A , B). 또한, 대부분의 세포 표현 심장 분 T, 분, 그리고 심장 α-actinin으로 갭 정션 마커 Connexin 43 하루 14 다음 변환 (<strong c...

Watch this Scientific Journal Video about Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes at JoVE.com

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

여기 선물이 overexpression Mef2c, GATA4, Hand2, Tbx5, 미르-1, 그리고 미르-133 (GHMT2m) TGF-β 신호 전달의 억제와 함께 통해 기능 cardiomyocytes에 기본 미 발달 섬유 아 세포를 재 설 정할 하는 강력한 방법. 우리의 프로토콜 생성 박동 cardiomyocytes 일찍 최대 60 %7 일 후 변환 효율.

프로 거리 신호 억제 유도 Cardiomyocytes에 마우스 미 발달 섬유 아 세포의 프로그래밍 요소 중재 Potentiates

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

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

신경의 분석을위한 성인 마우스 V-SVZ에서 세포의 안정적이고 효율적인 유전자 변형 줄기 세포 자율 및 비 자치 효과

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

연구

발생학

Improved Generation of Induced Cardiomyocytes Using a Polycistronic Construct Expressing Optimal Ratio of Gata4, Mef2c and Tbx5

Gata4, Mef2c 및 Tbx5의 최적 비율을 표현하는 폴리 시스 트론 구조를 사용하여 유도 된 심근의 향상 발전

Direct conversion of cardiac fibroblasts (CFs) into induced cardiomyocytes (iCMs) holds great potential for regenerative medicine by offering alternative ...

Protocol for the Direct Conversion of Murine Embryonic Fibroblasts into Trophoblast Stem Cells

영양막 줄기 세포로 쥐 배아 섬유 아세포의 직접적인 변환을위한 프로토콜

Trophoblast stem cells (TSCs) arise as a consequence of the first cell fate decision in mammalian development. They can be cultured in vitro, retaining ...

Reprogramming Mouse Embryonic Fibroblasts with Transcription Factors to Induce a Hemogenic Program

전사 인자와 마우스 배아 섬유 아 세포를 재 프로그램하는 것은 Hemogenic 프로그램을 유도하는

This protocol details the induction of a hemogenic program in mouse embryonic fibroblasts (MEFs) via overexpression of transcription factors (TFs). We ...

Stimulation of Notch Signaling in Mouse Osteoclast Precursors

마우스 파골 세포 전구체의 노치 신호 전달의 자극

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

Assessing Cardiomyocyte Subtypes Following Transcription Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts

마우스 배아 섬유 아 세포의 전사 인자 매개 재 프로그래밍에 따라 심근 서브 타입을 평가

Direct reprogramming of one cell type into another has recently emerged as a powerful paradigm for regenerative medicine, disease modeling, and lineage ...

Analysis of Retinoic Acid-induced Neural Differentiation of Mouse Embryonic Stem Cells in Two and Three-dimensional Embryoid Bodies

두와 입체 배아 체에서 마우스 배아 줄기 세포의 레티노 산에 의한 신경 분화 분석

Mouse embryonic stem cells (ESCs) isolated from the inner mass of the blastocyst (typically at day E3.5), can be used as in vitro model system for ...

Dissection and Culture of Mouse Embryonic Kidney

마우스 배아 신장의 해부 및 배양

The goal of this protocol is to describe a method for the dissection, isolation, and culture of mouse metanephric rudiments. 
During mammalian kidney ...

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

부상-유도 노화 및 골격 근육에서 프로그래밍을 Vivo에서 의 평가

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

Direct Lineage Reprogramming of Adult Mouse Fibroblast to Erythroid Progenitors

리니지 Reprogramming 성인 마우스 구와 Erythroid 창시자에의 직접

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