Name: isolation of preadipocytes from broiler chick embryos
Uploaded: 2022-08-04T13:00:00
Description: Watch this Scientific Journal Video about Isolation of Preadipocytes from Broiler Chick Embryos at JoVE.com

作者感谢UT农业研究和动物科学系支持和优化该协议。这项工作由美国农业部赠款资助。

所有动物程序均由田纳西大学机构动物护理和使用委员会批准。新鲜受精的商业肉鸡蛋（Cobb 500）是从当地的孵化场获得的。将卵在38°C下以60%的相对湿度孵育，直到胚胎第16-18天解剖（E16-E18）。从皮下（股骨）库收集脂肪组织。
1. 隔离和培养的准备
<ol><li>准备培养罩和仪器。<ol>
<li>在开始解剖之前，请在层流罩中设置一个工作区。通过用70%乙醇拭子对工作区域...

<ol>
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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+crude+oil/dispersant+mixture+and+dispersant+components+on+PPAR+&#947;+activity+in+vitro+and+in+vivo:+identification+of+dioctyl+sodium+sulfosuccinate+(DOSS;+CAS#+577-11-7)+as+a+probable+obesogen.">Effects of crude oil/dispersant mixture and dispersant components on PPAR &#947; activity in vitro and in vivo: identification of dioctyl sodium sulfosuccinate (DOSS; CAS# 577-11-7) as a probable obesogen.</a> Environmental Health Perspectives. 124, 112-119 (2016).</li><li>Hausman, D. B., Park, H. J., Hausman, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Adipose Tissue Protocols. , 201-219 (2008).</li><li>Lee, M. J., Wu, Y., Fried, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+modified+protocol+to+maximize+differentiation+of+human+preadipocytes+and+improve+metabolic+phenotypes.">A modified protocol to maximize differentiation of human preadipocytes and improve metabolic phenotypes.</a> Obesity. 20 (12), 2334-2340 (2012).</li><li>Church, C. D., Berry, R., Rodeheffer, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+study+of+adipocyte+precursors.">Isolation and study of adipocyte precursors.</a> Methods in Enzymology. 537, 31-46 (2014).</li><li>Akbar, N., Pinnick, K. E., Paget, D., Choudhury, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+characterization+of+human+adipocyte-derived+extracellular+vesicles+using+filtration+and+ultracentrifugation.">Isolation and characterization of human adipocyte-derived extracellular vesicles using filtration and ultracentrifugation.</a> Journal of Visualized Experiments. (170), e61979 (2021).</li><li>Perlman, D., Giuffre, N. A., Brindle, 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=Use+of+Fungizone+in+control+of+fungi+and+yeasts+in+tissue+culture.">Use of Fungizone in control of fungi and yeasts in tissue culture.</a> Proceedings of the Society for Experimental Biology and Medicine. 106 (4), 880-883 (1961).</li><li>Kuhlmann, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+prophylactic+use+of+antibiotics+in+cell+culture.">The prophylactic use of antibiotics in cell culture.</a> Cytotechnology. 19 (2), 95-105 (1995).</li><li>Yang, 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=Black+swimming+dots+in+cell+culture:+the+identity,+detection+method+and+judging+criteria.">Black swimming dots in cell culture: the identity, detection method and judging criteria.</a> bioRxiv. , 366906 (2018).</li><li>Freshney, R. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Culture of animal cells: a manual of basic technique and specialized applications. , 163-186 (2015).</li></ol>

尽管一些描述良好的方案已经报道了前脂肪细胞14，15，16，17的分离，但胚胎前脂肪细胞的分离已经优化，可用于肉鸡雏鸡早期脂肪生长和发育的功能分析。该方案产生具有高分化潜力的高生存能力的胚胎脂肪细胞祖细胞。此外，所提出的分离前脂肪细胞的程序不仅限于胚胎，还可以用于孵化后的雏?...

肥胖对成人和儿童都是一种全球健康威胁。超重或肥胖的儿童肥胖的可能性大约是成年的五倍，使他们患心血管疾病、糖尿病和许多其他合并症的风险显著增加。大约13.4%的美国2-5岁儿童患有肥胖症1，这表明积累多余身体脂肪的倾向可以在生命早期启动。由于非常不同的原因，过量脂肪组织的积累是肉鸡（肉型）鸡关注的问题。现代肉鸡非常高效，但仍然积累的脂质比生理上必需的多2，3.这种趋势在孵化后不久就开始了，并通过将其从肌肉生长中分配出来，有效地浪费了饲料，这是最昂贵的生产成分。因此，对于儿童和肉鸡，尽管出于非常不同的原因，但需要了解影响脂肪组织发育的因素，并确定在生命早期限制脂肪扩张的方法。
脂肪细胞形成于前脂肪细胞，脂肪组织来源的干细胞，经过分化以发展成熟的脂质储存脂肪细胞。因此， 体外 前脂肪细胞是肥胖研究的宝贵实验模型。这些从脂肪库的基质血管部分分离出来的细胞，可以提供对控制脂肪细胞分化和新陈代谢的分子途径的基本理解4，5。雏鸡胚胎是发育研究中一种有利的实验模型，因为按照所需的时间表培养卵子使实验操作更容易，因为它可以在没有母亲牺牲的情况下获得胚胎，以观察胚胎的一系列发育阶段。此外，相对于较大的动物模型，不需要复杂的外科手术和漫长的时间来获得胚胎。因此，雏鸡胚胎提供了从脂肪组织发育的最早阶段获得前脂肪细胞的机会。皮下脂肪组织在胚胎第12天（E12）左右的雏鸡中变得可见，作为位于大腿周围的明确定义的库。该库富含高度增殖的前脂肪细胞，这些前脂肪细胞在发育线索下积极经历分化以形成成熟的脂肪细胞6，7。脂肪分化的过程在鸡和人类之间是可比的。因此，从雏鸡胚胎中分离出的前脂肪细胞可以作为与人类和家禽相关的研究的两用模型。然而，随着细胞生长成成熟的脂肪细胞，前脂肪细胞的产量随着年龄的增长而下降5。
本方案优化了在肉鸡雏鸡胚胎中脂肪细胞分化和脂肪细胞肥大处于峰值的阶段（E16-E18）期间从脂肪组织中分离前脂肪细胞8。该过程可以评估发育 中的胚胎在卵中暴露的因素（例如母鸡饮食）对脂肪细胞发育和 离体脂肪电位的影响。它还可以测试各种操作（例如，缺氧、营养物质添加、药理激动剂和拮抗剂）对脂肪细胞祖细胞的脂肪生成或各种'组，代谢组，甲基组）的影响。作为脂肪形成最早阶段的表示，使用该方案获得的细胞是与家禽和人类相关的研究的宝贵模型。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1 mL Pipette</td>
			<td>Eppendorf</td>
			<td>Z683825</td>
			<td>Single Channel Pipette, 100 - 1000 &#181;L</td>
		</tr>
		<tr>
			<td>1 mL Pipette Tip</td>
			<td>Fisher Scientific</td>
			<td>02-707-402</td>
			<td></td>
		</tr>
		<tr>
			<td>100% Isopropanol</td>
			<td>Fisher Scientific</td>
			<td>A426P4</td>
			<td></td>
		</tr>
		<tr>
			<td>1x PBS</td>
			<td>Gibco</td>
			<td>10010023</td>
			<td></td>
		</tr>
		<tr>
			<td>25 mL Flask</td>
			<td>Pyrex</td>
			<td>4980-25</td>
			<td></td>
		</tr>
		<tr>
			<td>37% Formaldehyde</td>
			<td>Fisher Scientific</td>
			<td>F75P-1GAL</td>
			<td></td>
		</tr>
		<tr>
			<td>6-Well Plate</td>
			<td>Falcon</td>
			<td>353046</td>
			<td>Tissue Culture-treated</td>
		</tr>
		<tr>
			<td>96-Well Assay Plate</td>
			<td>Costar</td>
			<td>3632</td>
			<td></td>
		</tr>
		<tr>
			<td>96-Well Plate, Black Bottom</td>
			<td>Costar</td>
			<td>3603</td>
			<td>Tissue Culture-treated</td>
		</tr>
		<tr>
			<td>AdipoRed</td>
			<td>Lonza</td>
			<td>PT-7009</td>
			<td></td>
		</tr>
		<tr>
			<td>Amphotericin B</td>
			<td>Gibco</td>
			<td>15290026</td>
			<td></td>
		</tr>
		<tr>
			<td>Bench Top Wiper (Kimtechwiper)</td>
			<td>Kimberly-Clark</td>
			<td>34155</td>
			<td></td>
		</tr>
		<tr>
			<td>Betadine</td>
			<td>Up &#38; Up</td>
			<td>NDC 1167300334</td>
			<td>20% Working Solution</td>
		</tr>
		<tr>
			<td>Cell Counter</td>
			<td>Corning</td>
			<td>6749</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell Strainer, 40 &#181;m</td>
			<td>SPL</td>
			<td>93040</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifugaton</td>
			<td>Eppendorf</td>
			<td>5702</td>
			<td></td>
		</tr>
		<tr>
			<td>Chicken Serum</td>
			<td>Gibco</td>
			<td>16110082</td>
			<td></td>
		</tr>
		<tr>
			<td>Conical Centrifuge Tubes, 15 mL</td>
			<td>VWR</td>
			<td>10025-690</td>
			<td></td>
		</tr>
		<tr>
			<td>Conical Centrifuge Tubes, 50 mL</td>
			<td>Falcon</td>
			<td>352098</td>
			<td></td>
		</tr>
		<tr>
			<td>Cryovial</td>
			<td>Nunc</td>
			<td>343958</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved Forceps, 100 mm</td>
			<td>Roboz Surgical</td>
			<td>RS-5137</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved Surgical Scissors, 115 mm</td>
			<td>Roboz Surgical</td>
			<td>RS-6839</td>
			<td></td>
		</tr>
		<tr>
			<td>Distilled Water</td>
			<td>Millipore</td>
			<td>SYNSV0000</td>
			<td>Despensed as needed</td>
		</tr>
		<tr>
			<td>DMEM/F12</td>
			<td>HyClone</td>
			<td>SH30023.01</td>
			<td></td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>Sigma</td>
			<td>D2650</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Decon Labs</td>
			<td>2701</td>
			<td>70% Working Solution</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum (FBS)</td>
			<td>Gibco</td>
			<td>10437028</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescent Microscope</td>
			<td>Evos</td>
			<td>M7000</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescent Plate Reader</td>
			<td>Biotek</td>
			<td>Synergy H1</td>
			<td></td>
		</tr>
		<tr>
			<td>Foil</td>
			<td>Reynolds</td>
			<td></td>
			<td>Reynolds Wrap Heavy Duty Aluminum Foil, 125 SQ. FT.</td>
		</tr>
		<tr>
			<td>Freezing Container</td>
			<td>Thermo Scientific</td>
			<td>5100-0001</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelatin</td>
			<td>Millipore</td>
			<td>4055</td>
			<td>2% Working Solution</td>
		</tr>
		<tr>
			<td>Hematocytometer (Counting Chamber)</td>
			<td>Corning</td>
			<td>480200</td>
			<td>0.1 mm deep</td>
		</tr>
		<tr>
			<td>Incubator</td>
			<td>Fisher Scientific</td>
			<td>6845</td>
			<td></td>
		</tr>
		<tr>
			<td>Instrument Sterilizer</td>
			<td>VWR</td>
			<td>B1205</td>
			<td></td>
		</tr>
		<tr>
			<td>Linoleic Acid-Oleic Acid-Albumin</td>
			<td>Sigma</td>
			<td>L9655</td>
			<td>1x Working Solution</td>
		</tr>
		<tr>
			<td>Microscope</td>
			<td>Evos</td>
			<td>AMEX1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Multi-Channel Pipette</td>
			<td>Thermo Scientific</td>
			<td>4661070</td>
			<td>12-Channel Pipetters, 30 - 300 &#181;L</td>
		</tr>
		<tr>
			<td>Na2HPO4</td>
			<td>Sigma</td>
			<td>S-7907</td>
			<td></td>
		</tr>
		<tr>
			<td>NaH2PO4</td>
			<td>Sigma</td>
			<td>S-3139</td>
			<td></td>
		</tr>
		<tr>
			<td>NucBlue</td>
			<td>Invitrogen</td>
			<td>R37605</td>
			<td></td>
		</tr>
		<tr>
			<td>Oil Red O</td>
			<td>Sigma</td>
			<td>O-0625</td>
			<td></td>
		</tr>
		<tr>
			<td>Orbital Shaker</td>
			<td>IKA</td>
			<td>KS130BS1</td>
			<td></td>
		</tr>
		<tr>
			<td>Paper Towel</td>
			<td>Tork</td>
			<td>RK8002</td>
			<td></td>
		</tr>
		<tr>
			<td>Parafilm</td>
			<td>Parafilm M</td>
			<td>PM996</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin/Steptomycin (P/S)</td>
			<td>Gibco</td>
			<td>15140122</td>
			<td>1x Working Solution</td>
		</tr>
		<tr>
			<td>Petri dishes, 100 mm</td>
			<td>Falcon</td>
			<td>351029</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dishes, 60 mm</td>
			<td>Falcon</td>
			<td>351007</td>
			<td></td>
		</tr>
		<tr>
			<td>Plate Shaker</td>
			<td>VWR</td>
			<td>200</td>
			<td></td>
		</tr>
		<tr>
			<td>RBC Lysis Buffer</td>
			<td>Roche</td>
			<td>11814389001</td>
			<td></td>
		</tr>
		<tr>
			<td>Reagent Reservior</td>
			<td>VWR</td>
			<td>89094-680</td>
			<td></td>
		</tr>
		<tr>
			<td>Small Beaker, 100 mL</td>
			<td>Pyrex</td>
			<td>1000-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrophotometer Plate Reader</td>
			<td>Biotek</td>
			<td>Synergy H1</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile Gauze</td>
			<td>McKesson</td>
			<td>762703</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight Forceps, 120 mm</td>
			<td>Roboz Surgical</td>
			<td>RS-4960</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight Scissors, 140 mm</td>
			<td>Roboz Surgical</td>
			<td>RS-6762</td>
			<td></td>
		</tr>
		<tr>
			<td>T-25 Flask</td>
			<td>Corning</td>
			<td>430639</td>
			<td>Tissue Culture-treated</td>
		</tr>
		<tr>
			<td>Tissue Culture Incubator</td>
			<td>Thermo Scientific</td>
			<td>50144906</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue Strainer, 250 &#181;m</td>
			<td>Pierce</td>
			<td>87791</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypan Blue Stain</td>
			<td>Gibco</td>
			<td>15250061</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin</td>
			<td>Gibco</td>
			<td>15400054</td>
			<td>0.1% Working Solution</td>
		</tr>
		<tr>
			<td>Tweezers, 110 mm</td>
			<td>Roboz Surgical</td>
			<td>RS-5035</td>
			<td></td>
		</tr>
		<tr>
			<td>Type 1 Collagenase</td>
			<td>Gibco</td>
			<td>17100017</td>
			<td></td>
		</tr>
		<tr>
			<td>Water Bath</td>
			<td>Fisher Scientific</td>
			<td>15-462-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Whatman Grade 1 Filter Paper</td>
			<td>Whatman</td>
			<td>1001-110</td>
			<td></td>
		</tr>
	</tbody>
</table>

isolation of preadipocytes from broiler chick embryos

原代前脂肪细胞是了解控制脂肪细胞分化和代谢的分子途径的有价值的实验系统。鸡胚胎提供了从脂肪发育的最早阶段分离前脂肪细胞的机会。这种原代细胞可用于鉴定影响前脂肪细胞增殖和脂肪分化的因素，使其成为与儿童肥胖和控制家禽中多余脂肪沉积相关的研究的宝贵模型。产后脂肪组织的快速生长通过将其从肉鸡的肌肉生长中分配出来有效地浪费饲料。因此，了解脂肪组织发育最早阶段的方法可能为调节这种趋势提供线索，并确定在生命早期限制脂肪扩张的方法。本研究旨在开发一种有效的方法，用于从商业肉鸡（肉型）雏鸡胚胎的发育脂肪组织中分离出的前脂肪细胞的分离，初级培养和脂肪分化。该过程已经过优化，以产生具有高活力（约98%）的细胞，并增加了分化成成熟脂肪细胞的能力。这种简单的胚胎前脂肪细胞分离、培养和分化方法支持早期脂肪生长和发育的功能分析。

原代前脂肪细胞在形态上类似于成纤维细胞，具有不规则的星形形状和中央核（图2A-C）。细胞容易粘附在组织培养塑料上，并在附着后不久开始增殖。当在培养基中提供脂肪酸时，它们迅速分化并积累脂质液滴（图3D）。此处代表的分离物中报告的生存能力（98%，基于染料排除）是典型的。虽然细胞相当健壮，但在分离过程中...

Watch this Scientific Journal Video about Isolation of Preadipocytes from Broiler Chick Embryos at JoVE.com

Isolation of Preadipocytes from Broiler Chick Embryos

本方案描述了一种从肉鸡胚胎中的脂肪组织中分离前脂肪细胞的简单方法。该方法能够高产率、原代培养和前脂肪细胞的脂肪分化进行分离。油红O染色和脂质/DNA染色测量了分化培养基诱导的分离细胞的脂肪生成能力。

从肉鸡雏鸡胚胎中分离出前脂肪细胞

Primary preadipocytes are a valuable experimental system for understanding the molecular pathways that control adipocyte differentiation and metabolism. ...

原代前脂肪细胞是了解控制脂肪细胞代谢的分子途径的有价值的实验系统。鸡胚胎提供了从脂肪细胞发育的最早阶段分离前脂肪细胞的机会。该方法经过优化，产生具有高活力和增加的分化能力的细胞，可用于雏鸡早期脂肪生长发育胚胎的功能分析。鸡和人类之间的脂肪分化过程具有高度的可比性。因此，分离的前脂肪细胞可以作为与人类和家禽相关的研究的双重模型。开始用70%乙醇拭子胚胎体。然后为了防止过滤过程中的干扰，在消化后的后续步骤中用无菌纱布轻轻擦洗皮肤表面以除去羽毛。然后切掉两腿和腹部区域之间的皮肤，露出一对股骨脂肪库。用一只手用弯曲的镊子将皮肤固定在腿周围，轻轻去除股骨皮下脂肪。用另一只手。之后，使用弯曲的镊子轻轻地将长筒从腿部拉开。将组织转移到含有5毫升收集培养基的15毫升管中，并在脂肪垫的另一侧重复解剖。现在，将收集的脂肪垫转移到含有灭菌溶液的60毫米培养皿中，并通过旋转培养皿几次来短暂冲洗。然后通过将脂肪垫转移到含有PBS的60毫米培养皿中来冲洗灭菌溶液。轻轻旋转盘子，然后再次用PBS清洗。为了消化脂肪组织，将它们转移到含有预热酶溶液的15毫米管中。然后将一把长直剪刀浸入管中，最后将溶液中的脂肪组织切成尽可能小的碎片。将切碎的组织和酶溶液转移到25毫升高压灭菌的烧瓶中。用石蜡膜包裹烧瓶，并将烧瓶置于37摄氏度的轨道振荡器培养箱中进行消化。消化后，轻轻地上下移液以充分混合。然后通过移液将250微米的组织过滤器过滤到15毫升管中，以除去任何未消化的组织和碎片。现在，通过用4毫升生长培养基冲洗烧瓶来除去粘附在烧瓶上的细胞，并将细胞悬浮液过滤到相同的15毫升管中。接下来，通过再次移液生长培养基来冲洗过滤器，以除去过滤器中捕获的任何细胞。在室温下以300倍G离心5分钟，以沉淀细胞级分并吸出上清液而不干扰细胞沉淀。将沉淀轻轻重悬于一毫升红细胞裂解缓冲液中，通过移液充分混合并在室温下孵育五分钟。然后通过向含有细胞的管中加入五毫升生长培养基来稀释细胞，并通过移液轻轻混合它们。现在，将细胞移液到40微米的细胞过滤器上，并将它们过滤到一个新的50毫升管中。然后用另外五毫升的生长培养基冲洗过滤器，并通过在室温下以300倍G离心7分钟来沉淀细胞。小心地吸出上清液。通过移液将剩余的细胞沉淀重悬于一毫升生长培养基中 为了确定细胞密度和活力，通过移液将每个样品的10微升混合在台盼蓝中。将10微升混合物加载到血细胞计数器上并计数细胞。24、48和72小时后对前脂肪细胞的分析显示细胞形态和数量正常。前脂肪细胞的脂肪生成诱导显示脂质液滴的快速分化和积累。在分离过程中对前脂肪细胞的积极处理导致细胞损伤和低细胞数量。尽管采取了预防措施，仍可观察到微生物污染。用油红O染色脂质表明前脂肪细胞在脂肪诱导下迅速开始形成脂质液滴，并且随着时间的积累在24，48和72小时内观察到。使用脂质和DNA染色定量相对于细胞数的脂质积累。脂质积累和细胞增殖都随着时间的推移而增加。当组织部分被消化时间过长时，可以观察到低细胞数或受损细胞。因此，建议不要超过一个半小时的消化。分离的前脂肪细胞可用于基因表达研究。可以获得足够的RNA用于cDNA合成和后续的qPCR或RN酶。

Research

JoVE Journal

Developmental Biology

Optimized Ex-ovo Culturing of Chick Embryos to Advanced Stages of Development

Optimized Ex-ovo Culturing of Chick Embryos to Advanced Stages of Development

优化<em&gt;前OVO</em&gt;鸡胚培养，以发展的高级阶段

Research in anatomy, embryology, and developmental biology has largely relied on the use of model organisms. In order to study development in live embryos ...

科研

发育生物学

Forward Genetic Approach to Uncover Stress Resistance Genes in Mice &#8212; A High-throughput Screen in ES Cells

正向遗传学的方法来揭开抗逆基因小鼠 - 在ES细胞高通量筛选

Phenotype-driven genetic screens in mice is a powerful technique to uncover gene functions, but are often hampered by extremely high costs, which severely ...

Primary Endodermal Epithelial Cell Culture from the Yolk Sac Membrane of Japanese Quail Embryos

日本鹌鹑胚胎的卵黄囊膜主内胚上皮细胞培养

We established an endodermal epithelial cell culture model (EEC) for studying the function of certain enzymes and proteins in mediating nutrient ...

Isolation and Characterization of Single Cells from Zebrafish Embryos

分离和斑马鱼胚胎单细胞的表征

The zebrafish (Danio rerio) is a powerful model organism to study vertebrate development. Though many aspects of zebrafish embryonic development have been ...

Affinity Labeling Detection of Endogenous Receptors from Zebrafish Embryos

从斑马鱼胚胎内源性受体亲和标记检测

By combining the powers of Affinity Labeling and Immunoprecipitation (AFLIP), a technique for the detection of low abundance receptors in zebrafish ...

A Submerged Filter Paper Sandwich for Long-term Ex Ovo Time-lapse Imaging of Early Chick Embryos

A Submerged Filter Paper Sandwich for Long-term Ex Ovo Time-lapse Imaging of Early Chick Embryos

水下滤纸三明治长期<em&gt;防爆大毛</em&gt;早期鸡胚时间推移成像

Due to its availability, low cost, flat geometry, and transparency, the ex ovo chick embryo has become a major vertebrate animal model for the study of ...

Derivation of Stem Cell Lines from Mouse Preimplantation Embryos

从小鼠胚胎植入前胚胎干细胞系的推导

Mouse embryonic stem cell (mESC) derivation is the process by which pluripotent cell lines are established from preimplantation embryos. These lines ...

Preparation of Small RNA Libraries for Sequencing from Early Mouse Embryos

为早期小鼠胚胎测序准备小型RNA库

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

Generation of Na&#239;ve Blastoderm Explants from Zebrafish Embryos

从斑马鱼胚胎中产生幼稚的胚泡外植体

Due to their optical clarity and rapid development, zebrafish embryos are an excellent system for examining cell behaviors and developmental processes. ...