Name: production of extracellular matrix fibers via sacrificial hollow fiber membrane cell culture
Uploaded: 2019-02-02T13:00:00
Description: Watch this Scientific Journal Video about Production of Extracellular Matrix Fibers via Sacrificial Hollow Fiber Membrane Cell Culture at JoVE.com

该出版物中报告的研究得到了国家卫生研究院国家关节炎和肌肉骨骼及皮肤病研究所的支持, 该研究所的奖项号为 r15ar064481, 国家科学基金会 (cmmi-1404716), 以及阿肯色州生物科学研究所。

1. 使用牺牲中空纤维膜生产细胞外基质
注意: n-甲基-2-吡咯烷酮是一种刺激性溶剂和生殖毒物。接触 nmp 可能会对皮肤、眼睛、鼻子和喉咙造成刺激。在处理 nmp 时, 应使用耐溶剂的个人防护设备。nmp 的使用应在通风罩内进行。
<ol><li>中空纤维膜聚磺酸聚合物溶液的制备
	<ol>
<li>使用分析天平在称重船上称量70克聚磺酸颗粒 (分子量为 35kd)。</li>
		<li>将314毫升 (323.3 ...

<ol>
	<li>Cobb, W. S., Kercher, K. W., Heniford, B. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+argument+for+lightweight+polypropylene+mesh+in+hernia+repair.">The argument for lightweight polypropylene mesh in hernia repair.</a> Surgical Innovation. 12 (1), 63-69 (2005).</li><li>Morais, J. M., Papadimitrakopoulos, F., Burgess, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomaterials/Tissue+Interactions:+Possible+Solutions+to+Overcome+Foreign+Body+Response.">Biomaterials/Tissue Interactions: Possible Solutions to Overcome Foreign Body Response.</a> The AAPS Journal. 12 (2), 188-196 (2010).</li><li>Simon, P., 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=Early+failure+of+the+tissue+engineered+porcine+heart+valve+SYNERGRAFT&#174;+in+pediatric+patients.">Early failure of the tissue engineered porcine heart valve SYNERGRAFT&#174; in pediatric patients.</a> European Journal of Cardio-Thoracic Surgery. 23 (6), 1002-1006 (2003).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> FDA strengthens requirements for surgical mesh for the transvaginal repair of pelvic organ prolapse to address safety risks. , (2016).</li><li>Kartus, J., Movin, T., Karlsson, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Donor-site+morbidity+and+anterior+knee+problems+after+anterior+cruciate+ligament+reconstruction+using+autografts.">Donor-site morbidity and anterior knee problems after anterior cruciate ligament reconstruction using autografts.</a> Arthroscopy: The Journal of Arthroscopic &amp; Related Surgery. 17 (9), 971-980 (2001).</li><li>Lu, H., Hoshiba, T., Kawazoe, N., Chen, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autologous+extracellular+matrix+scaffolds+for+tissue+engineering.">Autologous extracellular matrix scaffolds for tissue engineering.</a> Biomaterials. 32 (10), 2489-2499 (2011).</li><li>Wolchok, J. C., Tresco, P. 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+isolation+of+cell+derived+extracellular+matrix+constructs+using+sacrificial+open-cell+foams.">The isolation of cell derived extracellular matrix constructs using sacrificial open-cell foams.</a> Biomaterials. 31 (36), 9595-9603 (2010).</li><li>Roberts, K., Schluns, J., Walker, A., Jones, J. D., Quinn, K. P., Hestekin, J., Wolchok, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+derived+extracellular+matrix+fibers+synthesized+using+sacrificial+hollow+fiber+membranes.">Cell derived extracellular matrix fibers synthesized using sacrificial hollow fiber membranes.</a> Biomedical Materials. 13 (1), (2017).</li><li>Hurd, S. A., Bhatti, N. M., Walker, A. M., Kasukonis, B. M., Wolchok, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+biological+scaffold+engineered+using+the+extracellular+matrix+secreted+by+skeletal+muscle+cells.">Development of a biological scaffold engineered using the extracellular matrix secreted by skeletal muscle cells.</a> Biomaterials. 49, 9-17 (2015).</li><li>Kasukonis, B. M., Kim, J. T., Washington, T. A., Wolchok, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+an+infusion+bioreactor+for+the+accelerated+preparation+of+decellularized+skeletal+muscle+scaffolds.">Development of an infusion bioreactor for the accelerated preparation of decellularized skeletal muscle scaffolds.</a> Biotechnology Progress. 32 (3), 745-755 (2016).</li><li>Murad, S., 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=Regulation+of+collagen+synthesis+by+ascorbic+acid.">Regulation of collagen synthesis by ascorbic acid.</a> Proceedings of the National Academy of Sciences of the United States of America. 78 (5), 2879-2882 (1981).</li><li>Zhang, Y., 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=Tissue-specific+extracellular+matrix+coatings+for+the+promotion+of+cell+proliferation+and+maintenance+of+cell+phenotype.">Tissue-specific extracellular matrix coatings for the promotion of cell proliferation and maintenance of cell phenotype.</a> Biomaterials. 30 (23-24), 4021-4028 (2009).</li><li>Feng, C. Y., Khulbe, K. C., Matsuura, T., Ismail, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+progresses+in+polymeric+hollow+fiber+membrane+preparation,+characterization+and+applications.">Recent progresses in polymeric hollow fiber membrane preparation, characterization and applications.</a> Separation and Purification Technology. 111, 43-71 (2013).</li><li>Domb, A. J., Kost, J., Wiseman, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of Biodegradable Polymers. , (1998).</li></ol>

所述工艺使使用干式湿纺系统铸造的中空纤维膜能够在体外生产散装 ecm 生物材料, 从而能够以低廉的价格批量生产膜以及标准的细胞培养设备。虽然本协议中制造的膜用于细胞培养, 但所述系统也可用于生产用于分离目的的膜, 孔径分布和中空纤维尺寸可通过变化进行调整纺丝机尺寸、所使用的聚合物、涂料和钻孔流量、取料速度和环境条件。13虽然该协议详细使用中空纤...

植入式手术支架是伤口修复的固定装置, 每年有超过100万个合成聚合物网格被植入全球, 仅用于腹壁修复1。然而, 植入后, 传统上用于制造这些支架的合成材料聚合物往往会引起异物反应, 导致炎症对植入物的功能有害, 组织结疤 2.此外, 由于主要的合成网状材料 (即聚丙烯) 并没有被人体明显改造, 它们一般适用于可以耐受疤痕的组织, 从而限制了它们在治疗方面的临床用途。具有高阶功能的组织, 如肌肉。虽然有许多外科网状产品已应用于临床上, 但最近制造商召回的合成手术网格和物种间组织植入物的并发症突出了最大限度地扩大植入物的重要性生物相容性, 促使 fda 收紧对手术网状企业 3,4的规定。从患者自身组织中提取的支架可降低这种免疫反应, 但可导致显著的供体-部位发病率 5。体外产生的细胞外基质 (ecm) 支架是一种可能的替代方法, 因为脱细胞化的 ecm 支架具有良好的生物相容性, 特别是在自体 ecm 植入物6的情况下。
由于患者组织用于自体植入的收获有限, 而且有可能阻碍供体部位的功能, 因此能够从人类细胞系的培养中, 或在可能的情况下, 从患者的细胞培养中生产体外的 ecm 支架。自己的细胞是一个有吸引力的选择。在体外制造大量 ecm 的主要挑战是这些难以捕获的分子的固存。在前面的工作中, 我们已经证明了 ecm 可以通过培养 ecm 分泌成纤维细胞在培养期后溶解的牺牲聚合物泡沫中产生 ecm, 从而产生可在植入7中脱珠化的 ecm, 8,9,10。由于泡沫中产生的 ecm 往往采用泡沫的内部结构, 中空纤维膜 (hfm) 被探索为生产 ecm 螺纹的牺牲支架。本文介绍了在成纤维细胞培养一段时间后, 负责细胞培养质量中空纤维膜实验室大规模制造和从同一细胞外基质纤维中提取的方法。这种静态培养方法很容易被含有标准哺乳动物细胞培养设备的实验室采用。这种方法产生的 ecm 可应用于各种临床应用。

<table>
	<tbody>
		<tr>
			<td>1/32 inch thick silicone rubber</td>
			<td>Grainger</td>
			<td>B01LXJULOM</td>
			<td></td>
		</tr>
		<tr>
			<td>20 mL Scintillation Vials, Borosilicate glass, Disposable - VWR</td>
			<td>VWR</td>
			<td>66022-004</td>
			<td>With attached white urea cap and cork foil liner</td>
		</tr>
		<tr>
			<td>3 inch by 1 inch microscopy slides</td>
			<td>VWR</td>
			<td>75799-268</td>
			<td></td>
		</tr>
		<tr>
			<td>4C refrigerator</td>
			<td>Thermo Fisher Scientific</td>
			<td>FRGG2304D</td>
			<td>Any commercial 4C refrigerator will suffice.</td>
		</tr>
		<tr>
			<td>50 mL tubes</td>
			<td>VWR</td>
			<td>21008-178</td>
			<td></td>
		</tr>
		<tr>
			<td>6-well cell culture plates</td>
			<td>VWR</td>
			<td>10062-892</td>
			<td>Alternative brands may be used</td>
		</tr>
		<tr>
			<td>Acetone</td>
			<td>VWR</td>
			<td>E646</td>
			<td>Alternative brands may be used</td>
		</tr>
		<tr>
			<td>Bore vessel</td>
			<td>McMaster-Carr</td>
			<td>89785K867</td>
			<td>6 ft 316 steel tubing</td>
		</tr>
		<tr>
			<td>Bovine Plasma Fibronectin</td>
			<td>Thermo Fisher Scientific</td>
			<td>33010018</td>
			<td>Comes as 1 mg of lyophilized protein</td>
		</tr>
		<tr>
			<td>CaCl2</td>
			<td>VWR/Amresco</td>
			<td>97062-590</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell Culture Incubator w/ CO2</td>
			<td></td>
			<td></td>
			<td>Any appropriate CO2-supplied mammalian cell incubator will suffice.</td>
		</tr>
		<tr>
			<td>Disposable Serological Pipets, Glass - Kimble Chase</td>
			<td>VWR</td>
			<td>14673-208</td>
			<td>Alternative brands may be used</td>
		</tr>
		<tr>
			<td>DMEM/F-12, HEPES</td>
			<td>Thermo Fisher Scientific</td>
			<td>11330032</td>
			<td>Warm in water bath at 37&#176;C for 30 minutes prior to use</td>
		</tr>
		<tr>
			<td>DNase I</td>
			<td>Sigma-Aldrich</td>
			<td>DN25-10MG</td>
			<td></td>
		</tr>
		<tr>
			<td>Dope vessel</td>
			<td>McMaster-Carr</td>
			<td>89785K867</td>
			<td>6 ft 316 steel tubing</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>VWR</td>
			<td>BDH1160</td>
			<td>Dilute to 70% for sterilization</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum, qualified, US origin - Gibco</td>
			<td>Thermo Fisher Scientific</td>
			<td>26140079</td>
			<td>Mix with growth media at 10% concentration (50mL in 500mL media)</td>
		</tr>
		<tr>
			<td>Four 1/4-inch to 1&#34; reducing unions</td>
			<td>Swagelok</td>
			<td>SS-1610-6-4</td>
			<td>One reducing union for each inlet and outlet of each vessel</td>
		</tr>
		<tr>
			<td>Freeze-dryer/lyophilizer</td>
			<td>Labconco</td>
			<td>117 (A65312906)</td>
			<td>Any lyophilizer will suffice.</td>
		</tr>
		<tr>
			<td>Hexagonal Antistatic Polystyrene Weighing Dishes - VWR</td>
			<td>VWR</td>
			<td>89106-752</td>
			<td>Any weigh boat will suffice</td>
		</tr>
		<tr>
			<td>Hollow fiber membrane immersion bath</td>
			<td></td>
			<td></td>
			<td>34L polypropylene tubs may be used or large bath containers can be fabricated from welded steel sheets</td>
		</tr>
		<tr>
			<td>Hollow Fiber Membrane Spinneret</td>
			<td>AEI</td>
			<td>http://www.aei-spinnerets.com/specifications.html</td>
			<td>Made to order. Inner diameter = 0.8 mm, outer diameter = 1.6 mm</td>
		</tr>
		<tr>
			<td>Hot plate/stirrer</td>
			<td>VWR</td>
			<td>97042-634</td>
			<td></td>
		</tr>
		<tr>
			<td>Human TGF-&#946;1</td>
			<td>PeproTech</td>
			<td>100-21</td>
			<td></td>
		</tr>
		<tr>
			<td>L-Ascorbic acid</td>
			<td>Sigma-Aldrich</td>
			<td>A4544-25G</td>
			<td></td>
		</tr>
		<tr>
			<td>L-Ascorbic acid 2-phosphate</td>
			<td>Sigma-Aldrich</td>
			<td>A8960-5G</td>
			<td></td>
		</tr>
		<tr>
			<td>L-glutamine (200 mM) - Gibco</td>
			<td>Thermo Fisher Scientific</td>
			<td>25030081</td>
			<td>Mix with growth media at 1% concentration (5mL in 500mL media)</td>
		</tr>
		<tr>
			<td>MgCl2</td>
			<td>VWR/Alfa Aesar</td>
			<td>AA12315-A1</td>
			<td></td>
		</tr>
		<tr>
			<td>Minus 80 Freezer</td>
			<td>Thermo Fisher Scientific</td>
			<td>UXF40086A</td>
			<td>Any commercial -80C freezer will suffice.</td>
		</tr>
		<tr>
			<td>N2 gas cylinders (two)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>NIH/3T3 cells</td>
			<td>ATCC</td>
			<td>CRL-1658</td>
			<td>Alternative fibrogenic cell lines may be used.</td>
		</tr>
		<tr>
			<td>N-methyl-2-pyrrolidone</td>
			<td>VWR</td>
			<td>BDH1141</td>
			<td>Alternative brands may be used</td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin Solution - Gibco</td>
			<td>Thermo Fisher Scientific</td>
			<td>15140122</td>
			<td>Mix with growth media at 0.1% concentration (0.5 mL in 500mL media)</td>
		</tr>
		<tr>
			<td>Polysulfone</td>
			<td>Sigma-Aldrich</td>
			<td>428302</td>
			<td>Any polysulfone with an average Mw of 35,000 daltons may be used</td>
		</tr>
		<tr>
			<td>Portable Pipet-Aid Pipetting Device - Drummond</td>
			<td>VWR</td>
			<td>53498-103</td>
			<td>Alternative brands may be used</td>
		</tr>
		<tr>
			<td>PTFE tubing (1/4-inch inner diameter)</td>
			<td>McMaster-Carr</td>
			<td>52315K24</td>
			<td>Alternative brands may be used.</td>
		</tr>
		<tr>
			<td>Rat skeletal muscle fibroblasts</td>
			<td></td>
			<td></td>
			<td>Independently isolated from rat skeletal muscle. Alternative fibrogenic cell lines may be used.</td>
		</tr>
		<tr>
			<td>RNase A</td>
			<td>Sigma-Aldrich</td>
			<td>R4642</td>
			<td></td>
		</tr>
		<tr>
			<td>Silicone sheet</td>
			<td>McMaster-Carr</td>
			<td>1460N28</td>
			<td></td>
		</tr>
		<tr>
			<td>Take-up motor</td>
			<td>Greartisan</td>
			<td>B071GTTSV3</td>
			<td>200 RPM DC Motor</td>
		</tr>
		<tr>
			<td>Tris HCl</td>
			<td>VWR/Amresco</td>
			<td>97063-756</td>
			<td></td>
		</tr>
		<tr>
			<td>Two needle valves</td>
			<td>Swagelok</td>
			<td>SS-1RS4</td>
			<td></td>
		</tr>
	</tbody>
</table>

production of extracellular matrix fibers via sacrificial hollow fiber membrane cell culture

来自细胞外基质 (ecm) 的工程支架使人们对医学产生了浓厚的兴趣, 因为它们在加速伤口闭合和愈合方面具有潜力。从体外纤维细胞培养中提取细胞外基质有可能从人类和潜在的患者特异性细胞系中产生 ecm, 从而最大限度地减少阻碍临床的异种表位的存在一些现有的 ecm 产品的成功。在体外生产适合植入的 ecm 中, 一个重大挑战是细胞培养的 ecm 生产通常产量相对较低。本文介绍了在牺牲中空纤维膜支架内培养的细胞生产 ecm 的方案。用成纤维细胞系在常规细胞培养基中培养中空纤维膜, 并在细胞培养后溶解, 产生连续的 ecm 螺纹。该方法制备的 ecm 纤维可脱珠化冻干, 适用于贮存和植入。

牺牲支架细胞外基质的成功生产取决于适当的支架制作、细胞培养和溶剂冲洗程序。中空纤维膜的制造是使用由市售部件组装的干射流湿纺系统进行的 (图 1), 该系统通过市售钢的环环挤出聚合物溶液纺丝器 (内径 = 0.8 毫米, 外径 = 1.6 毫米), 生成一个新生的聚合物溶液管, 在与水浴接触时沉淀成中空纤维膜。
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Watch this Scientific Journal Video about Production of Extracellular Matrix Fibers via Sacrificial Hollow Fiber Membrane Cell Culture at JoVE.com

Production of Extracellular Matrix Fibers via Sacrificial Hollow Fiber Membrane Cell Culture

该方案的目标是生产针对伤口修复的整体细胞外基质纤维, 作为再生支架植入物的一部分, 适合临床前评估。这些纤维是通过在中空纤维膜上培养成纤维细胞而产生的, 并通过膜的溶解提取。

牺牲纤维膜细胞培养生产细胞外基质纤维

Engineered scaffolds derived from extracellular matrix (ECM) have driven significant interest in medicine for their potential in expediting wound closure ...

该协议允许从培养细胞系生产散装细胞外基丝，允许研究细胞基质相互作用，以及研究这种生物材料对伤口修复的潜力。这些合成纤维可以避免针对合成材料的异物反应，同时仍然利用丰富的纺织制造技术，创造出广泛的编织植入材料。演示这个程序的将是卡桑德拉·里德，一个研究生，来自我的实验室。在细胞播种之前，在121摄氏度下将中空纤维膜在121摄氏度下进行30分钟，随后在PBS中用50毫升的牛血浆纤维素在PBS中用5%二氧化碳进行1小时的处理。在孵育结束时，使用无菌微剪刀将纤维切割到6厘米或以下，并使用装有21量针的1毫升注射器将10倍至第五纤维细胞直接植入中空纤维膜的亮度。将六种种子纤维放入直径为六英寸的培养皿中，并在37摄氏度的二氧化碳中短暂孵育种子纤维。五分钟后，将纤维从培养箱转移到一个新的六厘米培养皿中，含有纤维细胞培养基培养基，辅以L-抗坏血酸，L-抗坏血酸2磷酸盐，并在细胞培养箱中转换生长因子β1长达三周。在孵化结束时，使用钳子将培养的纤维转移到单个闪烁小瓶中，并倾斜每个小瓶，允许将多达五毫升的 N-甲基-2-pyrolidone 添加到小瓶的一侧。然后使用血清移液器从每根纤维中缓慢吸出N-甲基-2-丙酮，将纤维转移到新的闪烁小瓶中，以第二次浸入新鲜的N-甲基-2-丙酮。第三次浸入后，以类似的方式在脱离子水中冲洗产生的细胞外基体螺纹三次，将一块三英寸长、一英寸宽、1/23 英寸厚的硅橡胶与中央八×四毫米矩形模具放在标准的三×一英寸玻璃显微镜幻灯片上。高压灭菌后，将每个细胞外基质纤维并排放在八乘四毫米硅胶模具中，直到没有明显的开放空间。将模具放入50毫升锥形管中，将纤维冷冻在负80摄氏度，直到完全冻结。对于去细胞化，在1%的硫酸钠中孵育模具，在室温下用温和的搅拌机孵育24小时。第二天，用三毫升PBS洗涤，冲洗提取的细胞外基质，用新鲜准备好的DNA/RNA消化缓冲液填充模具，在4摄氏度下孵育6小时。在消化结束时，吸消化液，并在无菌PBS中冲洗模具三次，如证明。第三次洗涤后，在PBS中孵育10%青霉素-链霉素的脚手架在4摄氏度过夜，然后冻结在50毫升锥形管在负80摄氏度约1小时。当网状物完全冻结后，将去细胞化的细胞外网状在一夜之间冻干，或直到完全干燥，然后将冻干的脚手架转移到生物安全罩内的无菌容器中，并在四摄氏度下储存，直到使用。这里展示了使用本协议制造的聚糖中空纤维膜的横向横截面，展示了不对称膜的手指形毛孔外层和内层特征。在成纤维细胞播种和培养后，N-甲基-2-丙酮冲洗被证明，半透明线程的细胞外基质生产。在整个三周的培养期内，产生细胞的细胞外基质在中空纤维膜内仍然可行。提取的矩阵在水合时具有半透明外观，呈现非白色颜色和纤维外观，在网格装配和冻干时具有粗纵向对齐。最关键的一步是在N-甲基-2-丙酮溶剂中培养的空心纤维膜的溶解过程，以提取细胞外基质。该方法产生的材料可用于制造组织工程植入物，用于研究临床前研究中组织的修复。该协议中使用的 N-甲基-2-丙酮是皮肤和眼睛刺激物，因此建议您使用亚硝基手套、护目镜、实验室外套处理该化学品，并在烟罩内处理。

Research

JoVE Journal

Bioengineering

Non-contact, Label-free Monitoring of Cells and Extracellular Matrix using Raman Spectroscopy

非接触，无标签使用拉曼光谱研究细胞和细胞外基质的监测

Non-destructive, non-contact and label-free technologies to monitor cell and tissue cultures are needed in the field of biomedical research.1-5 However, ...

科研

生物工程

Epithelial Cell Repopulation and Preparation of Rodent Extracellular Matrix Scaffolds for Renal Tissue Development

上皮细胞复育和肾组织的发展准备啮齿动物细胞外基质的支架

This protocol details the generation of acellular, yet biofunctional, renal extracellular matrix (ECM) scaffolds that are useful as small-scale model ...

Hollow Fiber Bioreactors for In Vivo-like Mammalian Tissue Culture

Hollow Fiber Bioreactors for In Vivo-like Mammalian Tissue Culture

中空纤维生物反应器的<em&gt;在体内</em&gt;般的哺乳动物组织培养

Tissue culture has been used for over 100 years to study cells and responses ex vivo. The convention of this technique is the growth of anchorage ...

Engineering Three-dimensional Epithelial Tissues Embedded within Extracellular Matrix

工程三维上皮组织细胞外基质中嵌入

The architecture of branched organs such as the lungs, kidneys, and mammary glands arises through the developmental process of branching morphogenesis, ...

Preparation of Extracellular Matrix Protein Fibers for Brillouin Spectroscopy

细胞外基质蛋白纤维的布里渊谱的制备

Brillouin spectroscopy is an emerging technique in the biomedical field. It probes the mechanical properties of a sample through the interaction of ...

Tunable Hydrogels from Pulmonary Extracellular Matrix for 3D Cell Culture

可调水凝胶由肺细胞外基质的三维细胞培养

Here we present a method for establishing multiple component cell culture hydrogels for in vitro lung cell culture. Beginning with healthy en bloc lung ...

Fabrication of Extracellular Matrix-derived Foams and Microcarriers as Tissue-specific Cell Culture and Delivery Platforms

细胞外基质衍生的泡沫和微载体如组织特异性细胞培养和交付平台的制造

Cell function is mediated by interactions with the extracellular matrix (ECM), which has complex tissue-specific composition and architecture. The focus ...

Fabricating a Kidney Cortex Extracellular Matrix-Derived Hydrogel

肾皮质细胞外基质衍生水凝胶的制备

Extracellular matrix (ECM) provides important biophysical and biochemical cues to maintain tissue homeostasis. Current synthetic hydrogels offer robust ...

Pancreatic Tissue-Derived Extracellular Matrix Bioink for Printing 3D Cell-Laden Pancreatic Tissue Constructs

用于打印 3D 细胞-拉丹胰腺组织构造的胰腺组织衍生细胞外基质生物墨水

The transplantation of pancreatic islets is a promising treatment for patients who suffer from type 1 diabetes accompanied by hypoglycemia and secondary ...

A Human 3D Extracellular Matrix-Adipocyte Culture Model for Studying Matrix-Cell Metabolic Crosstalk

用于研究基质细胞代谢串扰的人类3D细胞外基质-脂肪培养模型

The extracellular matrix (ECM) plays a central role in regulating tissue homeostasis, engaging in crosstalk with cells and regulating multiple aspects of ...