作者要感谢：弗兰克博士姜大卫林博士，伯纳德Yurke博士和乌代Chippada博士为他们开发的DNA凝胶技术的贡献;诺雷尔Hadzimichalis博士，斯密特·沙阿，金佰利彼得曼，罗伯特·阿特尔的意见和这份手稿的编辑;资金来源包括新泽西州委员会对脊髓损伤研究（批准＃07A-019-SCR1，NAL）和新泽西州神经科学研究所（MLP）;与组织工程的出版商，A部分为许可转载图2和图4和生物材料的许可转载如图3所示 。

<ol>
	<li>Balaban, N. Q., 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=Force+and+focal+adhesion+assembly+A+close+relationship+studied+using+elastic+micropatterned+substrates.">Force and focal adhesion assembly A close relationship studied using elastic micropatterned substrates.</a> Nat Cell Biol. 3 (5), 466-472 (2001).</li><li>Peppas Brannon-Peppas, L., Peppas, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+and+equilibrium+swelling+behaviour+of+ph-sensitive+hydrogels+containing+2-hydroxyethyl+methacrylate.">Dynamic and equilibrium swelling behaviour of ph-sensitive hydrogels containing 2-hydroxyethyl methacrylate.</a> Biomaterials. 11 (9), 635-644 (1990).</li><li>Charati, M. B., Ifkovits, J. L., Burdick, J. A., Linhardt, J. G., Kiick, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrophilic+elastomeric+biomaterials+based+on+resilin-like+polypeptides.">Hydrophilic elastomeric biomaterials based on resilin-like polypeptides.</a> Soft Matter. 5 (18), 3412-3416 (2009).</li><li>Davis, K. A., Burke, K. A., Mather, P. T., Henderson, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+cell+behavior+on+shape+memory+polymer+substrates.">Dynamic cell behavior on shape memory polymer substrates.</a> Biomaterials. 32 (9), 2285-2293 (2011).</li><li>Dembo, M., Wang, Y. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stresses+at+the+cell-to-substrate+interface+during+locomotion+of+fibroblasts.">Stresses at the cell-to-substrate interface during locomotion of fibroblasts.</a> Biophys J. 76 (4), 2307-2316 (1999).</li><li>Gray, D. S., Tien, J., Chen, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repositioning+of+cells+by+mechanotaxis+on+surfaces+with+micropatterned+youngs+modulus.">Repositioning of cells by mechanotaxis on surfaces with micropatterned youngs modulus.</a> J Biomed Mater Res A. 66 (3), 605-614 (2003).</li><li>Homma, M., Seida, Y., Nakano, Y. <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+ions+on+the+dynamic+behavior+of+an+electrodriven+ionic+polymer+hydrogel+membrane.">Effect of ions on the dynamic behavior of an electrodriven ionic polymer hydrogel membrane.</a> Journal of applied Polymer Science. 82 (1), 76-80 (2001).</li><li>Horkay, F., Tasaki, I., Basser, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Osmotic+swelling+of+polyacrylate+hydrogels+in+physiological+salt+solutions.">Osmotic swelling of polyacrylate hydrogels in physiological salt solutions.</a> Biomacromolecules. 1 (1), 84-90 (2000).</li><li>Jiang, F. X., Yurke, B., Firestein, B. L., Langrana, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurite+outgrowth+on+a+DNA+crosslinked+hydrogel+with+tunable+stiffnesses.">Neurite outgrowth on a DNA crosslinked hydrogel with tunable stiffnesses.</a> Ann Biomed Eng. 36 (9), 1565-1579 (2008).</li><li>Jiang, F. X., Yurke, B., Schloss, R. S., Firestein, B. L., Langrana, N. A. <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+dynamic+stiffness+of+the+substrates+on+neurite+outgrowth+by+using+a+DNA-crosslinked+hydrogel.">Effect of dynamic stiffness of the substrates on neurite outgrowth by using a DNA-crosslinked hydrogel.</a> Tissue Engineering Part A. 16 (6), 1873-1889 (2010).</li><li>Jiang, F. X., Yurke, B., Schloss, R. S., Firestein, B. L., Langrana, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationship+between+fibroblast+growth+and+the+dynamic+stiffnesses+of+a+DNA+crosslinked+hydrogel.">The relationship between fibroblast growth and the dynamic stiffnesses of a DNA crosslinked hydrogel.</a> Biomaterials. 31 (6), 1199-1212 (2010).</li><li>Kloxin, A. M., Tibbitt, M. W., Anseth, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+photodegradable+hydrogels+as+dynamically+tunable+cell+culture+platforms.">Synthesis of photodegradable hydrogels as dynamically tunable cell culture platforms.</a> Nat Protoc. 5 (12), 1867-1887 (2010).</li><li>Lin, D. C., Yurke, B., Langrana, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+properties+of+a+reversible,+DNA-crosslinked+polyacrylamide+hydrogel.">Mechanical properties of a reversible, DNA-crosslinked polyacrylamide hydrogel.</a> J Biomech Eng. 126 (1), 104-110 (2004).</li><li>Lin, D. C., Yurke, B., Langrana, N. 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+rigid+spherical+inclusions+in+young's+moduli+determination:+Application+to+DNA-crosslinked+gels.">Use of rigid spherical inclusions in young's moduli determination: Application to DNA-crosslinked gels.</a> J Biomech Eng. 127 (4), 571-579 (2005).</li><li>Luo, Y., Shoichet, 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=Light-activated+immobilization+of+biomolecules+to+agarose+hydrogels+for+controlled+cellular+response.">Light-activated immobilization of biomolecules to agarose hydrogels for controlled cellular response.</a> Biomacromolecules. 5 (6), 2315-2323 (2004).</li><li>Marklein, R. A., Burdick, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatially+controlled+hydrogel+mechanics+to+modulate+stem+cell+interactions.">Spatially controlled hydrogel mechanics to modulate stem cell interactions.</a> Soft Matter. 6 (1), 136-143 (2010).</li><li>Pelham Jr, R. J., Wang, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+locomotion+and+focal+adhesions+are+regulated+by+substrate+flexibility.">Cell locomotion and focal adhesions are regulated by substrate flexibility.</a> Proc Natl Acad Sci U S A. 94 (25), 13661-13665 (1997).</li><li>Previtera, M. L., Chippada, U., Schloss, R. S., Yurke, B., Langrana, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+properties+of+DNA-crosslinked+polyacrylamide+hydrogels+with+increasing+crosslinker+density.">Mechanical properties of DNA-crosslinked polyacrylamide hydrogels with increasing crosslinker density.</a> BioResearch Open Access. 1 (5), 256-259 (2012).</li><li>Previtera, M. L., Langhammer, C. G., Firestein, B. L. <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+substrate+stiffness+and+cell+density+on+primary+hippocampal+cultures.">Effects of substrate stiffness and cell density on primary hippocampal cultures.</a> J Biosci Bioeng. 110 (4), 459-470 (2010).</li><li>Previtera, M. L., Langhammer, C. G., Langrana, N. A., Firestein, B. L. <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+dendrite+arborization+by+substrate+stiffness+is+mediated+by+glutamate+receptors.">Regulation of dendrite arborization by substrate stiffness is mediated by glutamate receptors.</a> Ann Biomed Eng. 38 (12), 3733-3743 (2010).</li><li>Previtera, M. L., Trout, K. L., Verma, D., Chippada, U., Schloss, R. S., Langrana, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fibroblast+morphology+on+dynamic+softening+of+hydrogels.">Fibroblast morphology on dynamic softening of hydrogels.</a> Ann Biomed Eng. 40 (5), 1061-1072 (2012).</li><li>Saxena, T., Gilbert, J., Stelzner, D., Hasenwinkel, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+characterization+of+the+injured+spinal+cord+after+lateral+spinal+hemisection+injury+in+the+rat.">Mechanical characterization of the injured spinal cord after lateral spinal hemisection injury in the rat.</a> J Neurotrauma. 29 (9), 1747-1757 (2012).</li><li>Sundararaghavan, H. G., Monteiro, G. A., Firestein, B. L., Shreiber, D. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurite+growth+in+3d+collagen+gels+with+gradients+of+mechanical+properties.">Neurite growth in 3d collagen gels with gradients of mechanical properties.</a> Biotechnol Bioeng. 102 (2), 632-643 (2009).</li><li>Wozniak, M. A., Chen, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanotransduction+in+development+A+growing+role+for+contractility.">Mechanotransduction in development A growing role for contractility.</a> Nat Rev Mol Cell Biol. 10 (1), 34-43 (2009).</li><li>Wuerfel, 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=Mr-elastography+reveals+degradation+of+tissue+integrity+in+multiple+sclerosis.">Mr-elastography reveals degradation of tissue integrity in multiple sclerosis.</a> Neuroimage. 49 (3), 2520-2525 (2012).</li><li>Zaari, N., Rajagopalan, P., Kim, S. K., Engler, A. J., Wong, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photopolymerization+in+microfluidic+gradient+generators+Microscale+control+of+substrate+compliance+to+manipulate+cell+response.">Photopolymerization in microfluidic gradient generators Microscale control of substrate compliance to manipulate cell response.</a> Adv Mater. 16 (23-24), 2133-2137 (2004).</li></ol>

作者什么都没有透露。

DNA凝胶的能力来软化或硬化之前和之后细胞粘附使得他们研究的动态组织硬度在细胞功能中的作用的理想模型。所有三种设计中采用了机械和生物学研究。然而，所有三个设计具有各种交联百分数类似弹性，表明交联长度不影响DNA的凝胶弹性（ 表2）。与此相反，丙烯酰胺浓度影响的弹性。这些设计可以不同交联动力学自凝胶破裂的倾向，或凝胶的倾向，在媒体或者缓冲溶解，随增?...

静态和动态的基材是两类生物材料的开发，以研究组织的弹性或刚性对细胞功能的影响的。静态基板无法它们被制造之后和/或一旦细胞被镀改变其物理性质。聚丙烯酰胺（PA）的凝胶进行合成的力学生物学研究5,17的被所述第一二维的，静态的基板。 PA凝胶是易于制备，价廉，通用性，并且可以与宽范围的弹性模量来制造。虽然这些技术的优点使PA胶凝共同施加的基板，静基板并不表明细胞外基质（ECM）和周围的细胞环境在体内的动态特性。例如，细胞外基质经历刚度的改变如损伤，发展，或疾病的结果。因此，动态的衬底作为力学生物学研究仿组织基底模型青睐<sup&gt; 22,24,25。 许多合成的，天然的，二维的，三 ​​维的，静态的和动态的生物材料已经被开发，以模仿组织硬度1,3,6,16,23,26。一些动态基板需要加热，紫外线，电流，离子和pH值的变化，以改变其机械性能2,4,7,8,12,15,16，但这些刺激可以限制水凝胶的生物应用。 DNA的交联聚丙烯酰胺水凝胶（DNA凝胶）是动态的二维弹性基材。 DNA的交联允许进行DNA凝胶刚度通过加入单链DNA（ssDNA）的时间，空间，和可逆调制到媒体或缓冲9-11,13,14,18,21。不像其中施加于弹性调制刺激上述的动态凝胶，将DNA凝胶依赖于应用的ssDNA为弹性的改变的扩散。因此，上部凝胶表面，其中细胞生长，是调制的，因为速度ø第一区域˚F弹性调制取决于凝胶的厚度。 DNA凝胶相似，其PA凝胶同行，他们有一个聚丙烯酰胺的骨干，但是双丙烯酰胺交联被替换为交联的DNA（ 图1）组成。两个的ssDNAs（SA1和SA2）杂交与交联剂链（L2），以弥补凝胶的DNA交联。 SA1和SA2有两个包含Acrydite修饰的5&#39;末端进行有效的结合到PA的网络不同序列。用于制备凝胶，SA1和SA2的单独聚合成PA骨架，随后聚合的SA1和SA2一起混合。 L2，交联剂，将被添加到SA1和SA2混合物。在L2的碱基序列互补的两个SA1和SA2的序列和二级杂交与SA1加上SA2以形成DNA交联。最初，DNA琼脂糖凝胶的弹性是由两个二级浓度和交联（ 表1确定</强&gt;和2）。含有L2，SA1，SA2和相等化学计量的量的DNA的凝胶是硬的凝胶，因为SA1和SA2是100％的交联用L2（被指定为100％的凝胶）。对二级导致DNA交联的比例较低，因此较低的浓度，柔和的DNA凝胶。凝胶低至50％的交联的（指定为50％凝胶）已经构建9-11。 <img alt="图1" src="/files/ftp_upload/51323/51323fig1highres.jpg" /> 图1中的DNA凝胶的交联和uncrosslinking概略9-11,13,14,18,21步骤1：SA1（红色）和SA2（蓝色）分别聚合成聚丙烯酰胺主链（黑色）。聚合后，SA1和SA2的聚合溶液混合在一起。步骤2：L 2（绿色），并将与SA1加上SA2杂交以形成凝胶的交联。第3步：R2的杂交与T他立足点L2的。步骤4：R2的立足点杂交推动L2的从SA1和SA2的解压。 与PA的凝胶，DNA凝胶可以变硬和合成后软化。出于这个原因，生长在DNA凝胶的细胞可以进行动态刚度的改变。变硬细胞粘附凝胶，L2可以加入到低百分比的凝胶培养基中，以增加交联点的百分比。软化细胞粘附凝胶，L2可以除去以减少交联点10,13,21的百分比。 L2具有在3&#39;端附加的立足点序列以允许L2从SA1和SA2中（ 表1）uncrosslink。去除L2的被逆转链称为R2的杂交来实现的。 R2是互补的L2的全长，并与L2立足点第一杂交。立足点杂交推动L2的从SA1和SA2的解压，这消除了交联，降低了凝胶的硬度。 在这份报告中，并视频，一步接一步提供了用于加固和软化的DNA凝胶的制备中说明。而100％和80％的凝胶制剂进行了描述，此协议可定制，以创建其他的最初和最后的交联百分比的DNA的凝胶。在一般情况下，100％和80％凝胶剂的制备，固定于盖玻片上，官能化的，并且接种有细胞。 L2加至80％凝胶介质和R2加至100％的凝胶剂，电镀后48小时的媒体。加L2的媒体变硬80％凝胶，以100％的交联，而加入R2到媒体软化100％的凝胶，以80％的交联。硬化的凝胶被指定为80→100％的凝胶和软凝胶中指定的文本100→80％的凝胶。为控制或静态凝胶，单链DNA组成的Ts或由于被传递到另一组的100％和80％的凝胶。至少两天下列弹性调制后，细胞可以进行处理和分析。 <table border="0" cellpadding="0" cellspacing="0" fo:kEEP-together.within页=“总是”&gt; <tbody align="center"><tr><td rowspan="2"></td><td rowspan="2"> DNA交联 </td><td rowspan="2"> ＃基地 </td><td> 序列（ 立足点 ） </td><td rowspan="2"> 修改 </td><td rowspan="2"> 熔化温度（Tm值，℃） </td><td rowspan="2"> 评论 </td></tr><tr><td> 5&#39;→3&#39; </td></tr><tr><td rowspan="4"> 设计1 </td><td> SA1 </td><td> 10 </td><td> GCA CCT TTGç </td><td> 5&#39;Acrydite </td><td> 34.9 </td><td></td></tr><tr><td> SA2 </td><td> 10 </td><td> GTC AGA ATGá </td><td> 5&#39;Acrydite </td><td> 23.6 </td><td></td></tr><tr><td> L2 </td><td> 30 </td><td> TCA TTC TGAç 气相色谱AAA级转肽酶的GC 摹CTA CAC得志 </td><td></td><td> 56 </td><td>10 bp的立足点序列包括在内。 </td></tr><tr><td> R2 </td><td> 30 </td><td> CAA GTG豪CGC ACC TTT GCG TCA GAA TGA </td><td></td><td></td><td> R2是互补的二级</td></tr><tr><td rowspan="4"> 设计2 </td><td> SA1 </td><td> 14 </td><td> CGT GGC ATA GGA CT </td><td> 5&#39;Acrydite </td><td> 46.​​9 </td><td></td></tr><tr><td> SA2 </td><td> 14 </td><td> GTT TCC民航局TCA GA </td><td> 5&#39;Acrydite </td><td> 40.2 </td><td></td></tr><tr><td> L2 </td><td> 40 </td><td> TCT GAT TGG GAA交流 教学专业议会注册税务师TGC CAC摹 GT交铁通卡特彼勒C </td><td></td><td> 65.9 </td><td>一个12碱基序列的立足点是包括在内。 </td></tr><tr><td> R2 </td><td> 40 </td><td> GAT GAA GGT AAC CGT GGC ATA GGA CTG TTT CCC AAT CAGá </td><td></td><td> 65.9 </td><td> R2是互补的二级</td></tr><tr><td rowspan="3"> DESIGñ3 </td><td> SA1 </td><td> 20 </td><td> ACG GAG GTG达GCA ATG TC </td><td> 5&#39;Acrydite </td><td> 55 </td><td></td></tr><tr><td> SA2 </td><td> 20 </td><td> 猫GCT TAG GGA CGA CTG GA </td><td> 5&#39;Acrydite </td><td> 56.6 </td><td></td></tr><tr><td> L2 </td><td> 40 </td><td> 台泥AGT CGT CCC认证TAA贲门癌的TG 摹ACA TTG猫ACA建CCG牛逼 </td><td></td><td> 68.8 </td><td>立足点不包括在内。 </td></tr><tr><td rowspan="2"> 控制 </td><td rowspan="2">控制</td><td rowspan="2"> 20-40 </td><td> AAA AAA（ 等 ），或</td><td rowspan="2"></td><td rowspan="2"></td><td rowspan="2"></td></tr><tr><td> TTT TTT（ 等等 ） </td></tr></tbody></table> 对单链DNA 9-11,13,14,18,21。细胞和机械的研究中采用了多种ð 表1中相应序列ifferent交联的设计，产生了一系列的静态和动态力学性能的DNA凝胶。调制的交联点的设计参数是碱基序列和序列长度或交联的长度。粗体和斜体字体说明SA1和L2之间和SA2和L2，分别之间的碱基配对。 <table border="0" cellpadding="0" cellspacing="0" fo:keep-together.within-page="always"><tbody align="center"><tr><td rowspan="2"></td><td colspan="8"> 设计 </td></tr><tr><td colspan="3"> 1 </td><td colspan="3"> 2 </td><td colspan="2"> 3 </td></tr><tr><td> 丙烯酰胺浓度（％） </td><td colspan="3"> 10 </td><td colspan="3"> 10 </td><td> 10 </td><td> 4 </td></tr><tr><td> SA1加上SA2杂交到二级（％交联） </td><td> 50 </td><td> 80 </td><td> 100 </td><td> 50 </td><td> 80 </td><td> 100 </td><td> 100 </td><td> 100 </td></tr><tr><td> &lt;STRONG&gt;弹性（千帕，平均值±标准差） </td><td> 6.6±0.6 </td><td> 17.1±0.8 </td><td> 29.8±2.5 </td><td> 5.85±0.62 </td><td> 12.67±1.33 </td><td> 22.88±2.77 </td><td> 25.2±0.5 </td><td> 10.4±0.6 </td></tr></tbody></table> 表2 DNA的凝胶9-11,13,14,18,21。丙烯酰胺浓度，交联百分比，和交联长度的杨氏模量（E）的可调制中的DNA凝胶。设计1，2，和3具有20，28和40碱基的交联​​长度，分别为。 100％的凝胶对于所有的设计也有类似的弹性模量表示的交联长度，不影响凝胶的弹性。但是，不同的丙烯酰胺浓度改变DNA的凝胶弹性。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>ssDNA</td>
			<td>Integrated DNA Technologies (Coralville, Iowa) idtdna.com</td>
			<td></td>
			<td>Do not vortex ssDNA. Gentle invert the vial and/or pipette solution to mix.</td>
		</tr>
		<tr>
			<td>PBS with calcium and magnesium</td>
			<td></td>
			<td></td>
			<td>Any brand.</td>
		</tr>
		<tr>
			<td>100X Tris-EDTA buffer (TE buffer)</td>
			<td>Sigma-Aldrich (St. Loius, MO) sigmaldrich.com</td>
			<td>T9285</td>
			<td></td>
		</tr>
		<tr>
			<td>10X Tris-Borate-EDTA buffer (TBE buffer)</td>
			<td>Sigma-Aldrich (St. Loius, MO)</td>
			<td>93290</td>
			<td>TBE is a reproductive toxin.</td>
		</tr>
		<tr>
			<td>40% Acrylamide solution</td>
			<td>Fisher Scientific (Pittsburg, PA)</td>
			<td>BP14021</td>
			<td>Acrylamide is a toxin.</td>
		</tr>
		<tr>
			<td>Ammonium persulfate (APS)</td>
			<td>Sigma-Aldrich (St. Loius, MO)</td>
			<td>A3678</td>
			<td>Prepare a 2% solution in TE buffer. APS is a toxin and irratant.</td>
		</tr>
		<tr>
			<td>Tetramethylethylenediamine (TEMED)</td>
			<td>Sigma-Aldrich (St. Loius, MO)</td>
			<td>T9281</td>
			<td>Prepare a 20% solution in TE buffer. TEMED is flammable, a corrosive, and a toxin.</td>
		</tr>
		<tr>
			<td rowspan="2">12-mm diameter round coverglass</td>
			<td rowspan="2">Fisher Scientific (Pittsburg, PA) fishersci.com</td>
			<td rowspan="2">12-545-82</td>
			<td rowspan="2"></td>
		</tr>
		<tr>
			<td>Norland optical adhesive 72</td>
			<td>Norland Products (Cranbury, NJ) norlandprod.com</td>
			<td>NOA72</td>
			<td></td>
		</tr>
		<tr>
			<td rowspan="2">24-well tissue culture plate</td>
			<td rowspan="2"></td>
			<td rowspan="2"></td>
			<td rowspan="2">Any brand.</td>
		</tr>
		<tr>
			<td rowspan="2">Microcentrifuge tubes</td>
			<td rowspan="2"></td>
			<td rowspan="2"></td>
			<td rowspan="2">Any brand.</td>
		</tr>
		<tr>
			<td>Sulfo-SANPAH</td>
			<td>ProteoChem or Thermo Fisher, (Rockland, IL) proteochem.com or thermofisher.com</td>
			<td>C111 or 22589</td>
			<td>Prepare a 0.315 mg/ml solution in water immediately before use. Dissolve at 37&#176;C and filter sterilze. It is normal to observe undisolved sulfo-SANPAH in the filter. Sulfo-SANPAH is light sensitive and, therefore, the solution should be protect from light until UV exposure.</td>
		</tr>
		<tr>
			<td>Poly-D-Lysine (PDL)</td>
			<td>Sigma-Aldrich (St. Loius, MO)</td>
			<td>P6407</td>
			<td>Prepare a 0.2 mg/ml solution in water and filter sterilize.</td>
		</tr>
		<tr>
			<td>Collagen Type I</td>
			<td>Affymetrix (Santa Clara, CA) affymetrix.com</td>
			<td>13813</td>
			<td>Prepare a 0.2 mg/ml solution in 0.2 N acetic acid. Solution needs to remain cold at all times to avoid polymerization. Acetic acid is a flammable, toxic, and corrosive.</td>
		</tr>
		<tr>
			<td>22 X 60 cover glass</td>
			<td>Fisher Scientific (Pittsburg, PA)</td>
			<td>12-544-G</td>
			<td></td>
		</tr>
		<tr>
			<td>Positive-displacement pipette</td>
			<td>Gilson, Inc (Middletown, WI) gilson.com</td>
			<td>F148504</td>
			<td></td>
		</tr>
		<tr>
			<td>Heat block</td>
			<td>Fisher Scientific (Pittsburg, PA)</td>
			<td>11-718</td>
			<td></td>
		</tr>
		<tr>
			<td>UV light source</td>
			<td></td>
			<td></td>
			<td>Place gels as close as possible to the UV light. UV light can cause skin or eye injury.</td>
		</tr>
		<tr>
			<td>Thermometer</td>
			<td></td>
			<td></td>
			<td>Any brand.</td>
		</tr>
		<tr>
			<td>Nitrogen gas</td>
			<td>GTS-Welco (Flemington, NJ) www.praxairmidatlantic.com/</td>
			<td>NI 5.0UH-R</td>
			<td></td>
		</tr>
	</tbody>
</table>

preparation of dna-crosslinked polyacrylamide hydrogels

力学生物学是一个新兴科学领域，解决物理线索中指导细胞的形态和功能中的关键作用。例如，组织的弹性对细胞功能的影响是力学生物学研究的一个主要领域，因为组织硬度调制与疾病，发展和损伤。静态仿组织材料，或材料，可以不改变刚度一旦细胞接种，都主要是用于研究组织刚度对细胞功能的影响。虽然从静态研究收集到的信息是有价值的，这些研究并不表示体内细胞微环境的动态性质。为了更好地解决对细胞功能的动态劲度的影响，我们开发了一种DNA的交联聚丙烯酰胺水凝胶系统（DNA凝胶）。不像其他动态基底，DNA的凝胶具有降低或制造之后增加刚度而不刺激的能力。 DNA凝胶组成的DNA crossl的被聚合成聚丙烯酰胺骨干油墨。加入并经由输送的单链DNA的去除交联允许的时间，空间，和凝胶弹性的可逆控制。我们已经表明在以前的报告中说动态DNA琼脂糖凝胶弹性的影响调制成纤维细胞和神经细胞的行为。在这份报告和视频，我们提供的示意图描述的DNA制备凝胶的DNA琼脂糖凝胶的交联机理，并一步一步的指示。

在此之前我们的研究，细胞外基质的相互作用，观察静态兼容生物材料或不可逆的单向动态基板。这些衬底不准确地反映细胞微环境的动态性质。我们的工作提供了研究的细胞外基质相互作用的软化和硬化动态生物材料更生理模型转移现有的技术范式。在以往的研究中，我们通过研究各种细胞的形态和表型上这些凝胶上分析细胞活力衬底的效果。在一个例子中，我们通过评价成纤维细胞形态学上?...

Watch this Scientific Journal Video about Preparation of DNA-crosslinked Polyacrylamide Hydrogels at JoVE.com

Preparation of DNA-crosslinked Polyacrylamide Hydrogels

我们的实验室已开发的DNA交联聚丙烯酰胺水凝胶，动态水凝胶体系，以更好地了解调节组织硬度对细胞功能的影响。这里，我们提供了示意图，说明和协议来制备这些凝胶。

DNA交联聚丙烯酰胺水凝胶的制备

Mechanobiology is an emerging scientific area that addresses the critical role of physical cues in directing cell morphology and function. For example, ...

preparation-of-dna-crosslinked-polyacrylamide-hydrogels

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14.3K 次观看.  JFK Medical Center. 我们的实验室已开发的DNA交联聚丙烯酰胺水凝胶，动态水凝胶体系，以更好地了解调节组织硬度对细胞功能的影响。这里，我们提供了示意图，说明和协议来制备这些凝胶。 