这项工作得到了欧洲联盟2020期 (赠款协议 645710) 和第七框架 (赠款协议612547和 319010) 方案的资助。作者希望感谢 Dr. Fotini Sourli-Chasioti 对活检程序的帮助。作者还感谢 Mr. Yiorgos Paterakis 在视频制作方面的宝贵帮助。

该研究符合《赫尔辛基宣言》规定的标准, 并获得萨利大学、运动科学系、#914; 我和 #959; 道德委员会. 
 1. 手术设备和消耗品的准备 <ol> <li> 将一个消毒的外科手术场放置在一个4轮手术的梅奥站上. </li>
	<li> 将下列耗材和手术设备放置在4轮手术的梅奥站上: 
	一双直操作剪刀 (15 厘米) 一手术刀 (No. 11) 一手术刀柄 一双弯曲剪刀 (14 厘米) 一只蚊子钳 一对科克镊子 一个外科镊子 聚维酮碘 (在无菌纱布) 一条缝合线 (4.0) 一双直剪刀 (11 厘米) 一针架 (15 厘米) 五无菌纱布 一胶粘剂无菌纱布 一10毫升注射器与消毒针 10mL 2% 因 (没有肾上腺素) 一消毒手术手套 </li> <li> 将以下材料放在单独的工作台上: 两个离心管 一管含有5毫升的10% 福尔马林 一个小容器, 其中含有50-100 毫升液氮在-190 和 #176;</li> </ol> 2。参加者的准备 <ol> <li> 在活检过程之前让参与者遵循8小时的快速 15 。
	<li> 让参加者在活检之前不要进行体育锻炼, 过度的压力, 酒精, 主动和被动吸烟72小时 16 , 17 . </li>
</ol> 3。执行皮下脂肪组织活检 <ol> <li> 将参与者置于仰卧位的手术床上. </li>
	<li> 使用聚维酮碘对切口区域进行消毒. </li>
	<li> 将10毫升的2% 因 (无肾上腺素) 缓慢而稳步地注入到选定区域中, 作为局部麻醉. </li>
	<li> 在该区域放置一个无菌手术场, 以覆盖参与者的身体, 不包括切口区域. </li>
	<li> 至少3-5 分钟后, 用一对镊子按压局部麻醉区域, 并询问参与者是否感到疼痛 19 以确保局部麻醉已实现. </li>
	<li> 将 No. 11 手术刀附在手术刀柄上. </li>
	<li> 创建切口 2-2. 5 厘米长度, 3-5 厘米到左边或右侧的肚脐. </li>
	<li> 用一对直15厘米的操作剪刀将皮下组织取出. </li>
	<li> 一旦脂肪组织显露, 用一对镊子和一对弯曲的14厘米剪刀捕捉它. </li>
	<li> 切割并除去大约1克脂肪组织. </li>
	<li> 组织切除后, 在切口处放置无菌纱布, 以达到止血效果. </li>
	<li> 当止血被证实时, 用14厘米针和皮肤用一双外科镊子捕捉4.0 缝合, 直到皮下组织可见. </li>
	<li> 将缝合针插入皮肤, 向皮下组织移动, 而皮肤和皮下紧密排列. </li>
	<li> 用缝合针, 沿着一个连续的曲折技术进入皮下组织, 直到从皮肤外部驱动。继续, 直到皮肤切口关闭. </li>
	<li> 外部绑住缝合线 </li>
	<li> 用生理盐水清洗切口面积, 并用无菌纱布覆盖. </li>
</ol> 4。组织的沉积 <ol> <li> 将收集的脂肪组织分离为三块: 150、350和500毫克. </li>
	<li> 将150和350毫克样品组织放置在离心管中. </li>
	<li> 将两个离心管浸入小容器中, 将液氮浸泡在-190 和 #176; c 直到在-80 和 #176 的冷冻库中最终沉积; c. 
	注: 预先使用无菌针头刺穿离心管中的一个小孔, 以避免在-190 和 #176 的液氮浸入离心管时发生杯状故障; C. </li>
	<li> 浸泡500毫克样品组织-用于组织学-一管含有5毫升10% 福尔马林. 
	注: 只有在手术过程中出血过多的情况下才使用蚊虫钳。在这种情况下, 捕获的船只与蚊子钳结扎和关闭, 直到止血达到. </li>
</ol>

<ol><li>Bruening, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Systematic+review%3A+comparative+effectiveness+of+core-needle+and+open+surgical+biopsy+to+diagnose+breast+lesions%5BTitle%5D)+AND+%22Ann+Intern+Med%22%5BJournal%5D)">Systematic review: comparative effectiveness of core-needle and open surgical biopsy to diagnose breast lesions.</a> Ann Intern Med. 152, 238-246 (2010).</li>
<li>Beynen, A. C., Katan, M. B. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/4025201">Rapid sampling and long-term storage of subcutaneous adipose-tissue biopsies for determination of fatty acid composition.</a> Am J Clin Nutr. 42, 317-322 (1985).</li>
<li>Wang, Y., et al. <a target="_blank" href="http://file.scirp.org/pdf/JDM20110300002_53765035.pdf">Muscle and adipose tissue biopsy in older adults with type 2 diabetes .</a> J Diabetes Mellitus. 1, 27-35 (2011).</li>
<li>Mutch, D. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Needle+and+surgical+biopsy+techniques+differentially+affect+adipose+tissue+gene+expression+profiles%5BTitle%5D)+AND+%22Am+J+Clin+Nutr%22%5BJournal%5D)">Needle and surgical biopsy techniques differentially affect adipose tissue gene expression profiles.</a> Am J Clin Nutr. 89, 51-57 (2009).</li>
<li>Ikeda, S., Sekijima, Y., Tojo, K., Koyama, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Diagnostic+value+of+abdominal+wall+fat+pad+biopsy+in+senile+systemic+amyloidosis%5BTitle%5D)+AND+%22Amyloid%22%5BJournal%5D)">Diagnostic value of abdominal wall fat pad biopsy in senile systemic amyloidosis.</a> Amyloid. 18, 211-215 (2011).</li>
<li>Alderete, T. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+novel+biopsy+method+to+increase+yield+of+subcutaneous+abdominal+adipose+tissue%5BTitle%5D)+AND+%22Int+J+Obes%22%5BJournal%5D)">A novel biopsy method to increase yield of subcutaneous abdominal adipose tissue.</a> Int J Obes. 39, 183-186 (2015).</li>
<li>Padoin, A. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+comparison+of+wedge+and+needle+hepatic+biopsy+in+open+bariatric+surgery%5BTitle%5D)+AND+%22Obes+Surg%22%5BJournal%5D)">A comparison of wedge and needle hepatic biopsy in open bariatric surgery.</a> Obes Surg. 16, 178-182 (2006).</li>
<li>Kral, J. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+surgical+treatment+of+the+metabolic+syndrome+on+liver+fibrosis+and+cirrhosis%5BTitle%5D)+AND+%22Surgery%22%5BJournal%5D)">Effects of surgical treatment of the metabolic syndrome on liver fibrosis and cirrhosis.</a> Surgery. 135, 48-58 (2004).</li>
<li>Fitzgerald, J. E., Malik, M., Ahmed, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+single-blind+controlled+study+of+electrocautery+and+ultrasonic+scalpel+smoke+plumes+in+laparoscopic+surgery%5BTitle%5D)+AND+%22Surg+Endosc%22%5BJournal%5D)">A single-blind controlled study of electrocautery and ultrasonic scalpel smoke plumes in laparoscopic surgery.</a> Surg Endosc. 26, 337-342 (2012).</li>
<li>Memon, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Surgical+Diathermy%5BTitle%5D)+AND+%22Br+J+Hosp+Med%22%5BJournal%5D)">Surgical Diathermy.</a> Br J Hosp Med. 52, (1994).</li>
<li>Tomita, Y., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/7027028">Mutagenecity of smoke condensates induced by CO-laser irradiation and electrocauterization.</a> Mutat Res. 89, 145-149 (1989).</li>
<li>Brandon, H., Young, L. <a target="_blank" href="https://www.surgimedics.com/Research%20Articles/Electrosurgical%20Plume/Characterization%20And%20Removal%20Of%20Electrosurgical%20Smoke.pdf">Characterisation and removal of electrosurgical smoke.</a> Surg Serv Manag. 3, 14-16 (1997).</li>
<li>Attner, P., Hemlin, C., Soderman, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ligasure+versus+diathermy+scissors+tonsillectomy%3A+A+controlled+randomized+study%5BTitle%5D)+AND+%22Acta+Otolaryngol%22%5BJournal%5D)">Ligasure versus diathermy scissors tonsillectomy: A controlled randomized study.</a> Acta Otolaryngol. 130, 1180-1184 (2010).</li>
<li>Ecker, T., Carvalho, A. L., Choe, J. H., Walosek, G., Preuss, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Hemostasis+in+thyroid+surgery%3A+harmonic+scalpel+versus+other+techniques--a+meta-analysis%5BTitle%5D)+AND+%22Otolaryngol+Head+Neck+Surg%22%5BJournal%5D)">Hemostasis in thyroid surgery: harmonic scalpel versus other techniques--a meta-analysis.</a> Otolaryngol Head Neck Surg. 143, 17-25 (2010).</li>
<li>Campbell, K. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+pilot+study+of+sampling+subcutaneous+adipose+tissue+to+examine+biomarkers+of+cancer+risk%5BTitle%5D)+AND+%22Cancer+Prev+Res%22%5BJournal%5D)">A pilot study of sampling subcutaneous adipose tissue to examine biomarkers of cancer risk.</a> Cancer Prev Res. 2, 37-42 (2009).</li>
<li>Jamurtas, A. Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+effects+of+a+single+bout+of+exercise+on+resting+energy+expenditure+and+respiratory+exchange+ratio%5BTitle%5D)+AND+%22Eur+J+Appl+Physiol%22%5BJournal%5D)">The effects of a single bout of exercise on resting energy expenditure and respiratory exchange ratio.</a> Eur J Appl Physiol. 92, 393-398 (2004).</li>
<li>Adriaens, P. E. M., Schoffelen, F. M. P., Westerterp, K. R. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/12908903">Intra-individual variation of basal metabolic rate and the inﬂuence of daily habitual physical activity before testing.</a> Brit J Nutri. 90, 419-423 (2003).</li>
<li>Latham, J. L., Martin, S. N. <a target="_blank" href="http://www.aafp.org/afp/2014/0615/p956.html">Infiltrative anesthesia in office practice.</a> Am Fam Phys. 89, 956-962 (2014).</li>
<li>Jurevic, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Plasma+levels+of+2%25+lidocaine+with+1%3A100%2C000+epinephrine+with+young+children+undergoing+dental+procedures%5BTitle%5D)+AND+%22Anesth+Prog%22%5BJournal%5D)">Plasma levels of 2% lidocaine with 1:100,000 epinephrine with young children undergoing dental procedures.</a> Anesth Prog. 45, 87-90 (1998).</li>
<li>Kale, S. S., Yende, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+Aging+on+Inflammation+and+Hemostasis+through+the+Continuum+of+Critical+Illness%5BTitle%5D)+AND+%22Aging+Dis%22%5BJournal%5D)">Effects of Aging on Inflammation and Hemostasis through the Continuum of Critical Illness.</a> Aging Dis. 2, 501-511 (2011).</li>
</ol>

作者声明他们没有竞争的金融利益。

本研究的目的是规范一项手术技术的皮下腹部脂肪组织活检, 进行局部麻醉下进行了 non-diathermy 方法的健康男性。透在手术室里常用;但是, 可能出现的副作用 (即,闪火和环境毒性)9和愈合中的延迟10 。透的这些有害影响会给操作人员和患者/参与者带来健康风险。因此, 有必要进行皮下脂肪组织活检的手术 non-diathermy 方法。
在手术后两?...

脂肪组织活检提供组织样本, 经分析后, 可以提供有关新陈代谢和疾病的机制的有见地的概述。例如, 它们用于乳腺癌肿瘤的鉴定1, 脂肪酸组成检查的2, 以及重要的代谢和荷尔蒙疾病通路的研究3。为了获得腹部区域的皮下脂肪组织活检, 研究人员和医生使用的是外科手术或 needle-based 技术。然而, 手术皮下脂肪活检可以提供组织样本, 可能提供一个更全面的概述复杂的生物机制 [例如,信使核糖核酸 (mRNA) 的表达, 组织学分析,等]4.它们以前已用于健康成人4 , 可在床边5中安全地执行。
needle-based 活检技术需要强烈的抽脂, 可能破坏组织的完整性, 它通常检索少量的组织 (100-500 毫克)6。这些组织标本可能不适合复杂的研究研究。此外, 与 needle-based 技术相比, 手术皮下脂肪活检可以提供组织样本, 可以绘制一个更完整的图片的几个炎症指数 (例如, b-和 T 细胞信号);细胞骨架调节;细胞通路;脂质、碳水化合物和氨基酸的代谢途径;和氧化磷酸化途径4。此外, needle-based 技术包括更高的风险, 血液细胞污染比外科手术一个4, 而提供了关于组织的纤维化区域的小信息4,7,8.
通常, 手术脂肪组织活检包括透治疗烧灼血管, 以防止过多出血9。透也被用于烧伤和破坏受疾病感染的组织和疣或组织的影响9。这项技术特别适用于神经外科和眼科手术9。
过度使用透可能导致坏死组织的形成, 导致感染和延迟愈合过程10。事实上, 在透9之后, 报告的副作用如闪光火和组织中的皮肤损害。此外, 有人担心透的外科烟雾对手术环境的毒性, 这可能会被参与者/病人和操作人员吸入9。事实上, 使用透可能会导致手术室空气的毒性等于吸烟六香烟的11和, 虽然手术室通常有良好的通风系统, 污染可以持续至少20分钟12.然而, 在进行扁桃体手术的患者中, 透方法显示出较少, 但显著, 术后疼痛13和出血的14比传统的结扎止血方法。总的来说, 关于健康个体脂肪组织活检后疼痛和出血率差异的证据几乎不存在。鉴于缺乏证据和透的副作用, 有必要对脂肪组织活检的 non-diathermy 方法进行标准化。因此, 本研究的目的是规范一项手术技术的皮下腹部脂肪组织活检, 进行局部麻醉下使用 non-diathermy 法的健康男性。此外, 我们概述了组织收集的程序, 以确定 mRNA 的表达和蛋白质浓度, 并进行组织学分析。

<table><tbody><tr><td>&amp;nbsp;Disinfected surgical field&amp;nbsp;</td><td>Hartmann</td><td>23516-00</td><td></td></tr><tr><td>Stainless Steel Instrument Stands (surgical Mayo)</td><td>Medical Device Depot Inc.</td><td>32000</td><td></td></tr><tr><td>Operating Scissors straight 15 cm</td><td>Karl Storz</td><td>791903</td><td></td></tr><tr><td>Scalpel No 11&amp;nbsp;</td><td>Swann-Morton</td><td>203</td><td>Bipolar, catalog number 429011</td></tr><tr><td>Scarpel handle No 3</td><td>Karl Storz</td><td>488090</td><td></td></tr><tr><td>Scissors curved 14 cm</td><td>Karl Storz</td><td>511514, 752918</td><td></td></tr><tr><td>Mosquito forceps</td><td>Karl Storz</td><td>535012</td><td></td></tr><tr><td>Tweezers Kocher&amp;nbsp;</td><td>Karl Storz</td><td>530416</td><td>Bipolar, catalog number 830316</td></tr><tr><td>Surgical tweezers</td><td>Karl Storz</td><td>793216</td><td>Bipolar, catalog number 831016</td></tr><tr><td>Povidone-iodine&amp;nbsp;</td><td>Various</td><td>Not applicable</td><td></td></tr><tr><td>Suture 4.0 Vicryl</td><td>Johnsons</td><td>V4970H&amp;nbsp;</td><td></td></tr><tr><td>Scissors straight 11 cm</td><td>Karl Storz</td><td>512511 DS</td><td></td></tr><tr><td>Needle holder 15 cm</td><td>Karl Storz</td><td>515515</td><td>Bipolar, catalog number 213015</td></tr><tr><td>&amp;nbsp;Sterile gauzes</td><td>Hartmann</td><td>3021</td><td></td></tr><tr><td>&amp;Alpha;dhesive sterile gauze&amp;nbsp;</td><td>Digas medical equipment</td><td>2889</td><td></td></tr><tr><td>10 mL syringe with disinfected needle</td><td>Alpha medical solutions</td><td>10310-55-0010</td><td></td></tr><tr><td>2%-xylocaine (no adrenaline)</td><td>Various</td><td>Not applicable</td><td></td></tr><tr><td>Sterile surgical gloves&amp;nbsp;</td><td>Sempermed</td><td>10001</td><td></td></tr><tr><td>Eppendorf tubes</td><td>Kartell</td><td>16283</td><td></td></tr><tr><td>10% formalin&amp;nbsp;</td><td>Alpha medical solutions</td><td>11016-30-2909</td><td></td></tr><tr><td>Tubes 120 mL</td><td>Digas medical equipment</td><td>9025</td><td></td></tr><tr><td>Liquid nitrogen at -190&amp;deg;C</td><td>Revival</td><td>Not applicable</td><td></td></tr></tbody></table>

a technique for subcutaneous abdominal adipose tissue biopsy via a non-diathermy method

脂肪组织活检提供组织样本, 经分析后, 可提供有关代谢和疾病机制的有见地的概述。为了获得腹部区域的皮下脂肪组织活检, 研究人员和医生使用的是外科手术或 needle-based 技术。然而, 手术皮下脂肪活检可以提供组织样本, 可以提供更全面的概述的复杂性生物指标的白色脂肪组织。通常, 手术脂肪组织活检包括透治疗烧灼血管, 以防止过度出血。然而, 在透后, 有报道称, 在组织中出现了闪光和皮肤损伤等副作用。因此, 我们的目的是规范一个外科腹部脂肪组织活检进行局部麻醉使用 non-diathermy 方法。我们进行了115皮下脂肪组织活检的健康男性使用 non-diathermy 腹部手术活检方法。我们的结果显示, 三例术后出血过多, 115 手术 (2.61%)。总之, 我们的标准化皮下腹部脂肪组织手术活检使用 non-diathermy 方法可以安全地应用于健康的人在床边, 最小的副作用。

参与者的特征在表 1中提供。切口区域的缝线在接下来的8-12 天内脱落。我们一共完成了115皮下脂肪组织活检使用 non-diathermy 腹部外科活检方法在健康成人 (年龄: 35.2 ±6.9 年; 身体质量索引: 27.3 ± 4.4; 和身体脂肪百分比: 27.0 ± 9.0%)。术后两天观察三例 (患病率: 2.61%) 后活检过度出血。三名经历过大出血的人比没有经历过任何副作用的人年长 (t(113)=-2.115, p ...

Watch this Scientific Journal Video about A Technique for Subcutaneous Abdominal Adipose Tissue Biopsy via a Non-diathermy Method at JoVE.com

A Technique for Subcutaneous Abdominal Adipose Tissue Biopsy via a Non-diathermy Method

我们用局部麻醉下的 non-diathermy 法对腹部脂肪组织进行了活检。三例术后大出血115手术 (2.61%) 发生。我们的结论是, 腹部脂肪组织手术活检使用 non-diathermy 的方法可以安全地应用于健康男性。

经非透法皮下腹脂肪组织活检技术

Adipose tissue biopsies offer tissue samples that, upon analysis, may provide insightful overviews of mechanisms relating to metabolism and disease. To ...

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15.0K 次观看.  University of Thessaly. 我们用局部麻醉下的 non-diathermy 法对腹部脂肪组织进行了活检。三例术后大出血115手术 (2.61%) 发生。我们的结论是, 腹部脂肪组织手术活检使用 non-diathermy 的方法可以安全地应用于健康男性。

视频: A Technique for Subcutaneous Abdominal Adipose Tissue Biopsy via a Non-diathermy Method

Performing and Processing FNA of Anterior Fat Pad for Amyloid

Historically, heart, liver, and kidney biopsies were performed to demonstrate amyloid deposits in amyloidosis. Since the clinical presentation of this disease is so variable and non-specific, the associated risks of these biopsies are too great for the diagnostic yield. Other sites that have a lower biopsy risk, such as skin or gingival, are also relatively invasive and expensive. In addition, these biopsies may not always have sufficient amyloid deposits to establish a diagnosis. Fat pad aspiration has demonstrated good clinical correlation with low cost and minimal morbidity. However, there are no standardized protocols for performing this procedure or processing the aspirated specimen, which leads to variable and nonreproducible results. The most frequently utilized modality for detecting amyloid in tissue is an apple-green birefringence on Congo red stained sections using a polarizing microscope. This technique requires cell block preparation of aspirated material. Unfortunately, patients presenting in early stage of amyloidosis have minimal amounts of amyloid which greatly reduces the sensitivity of Congo red stained cell block sections of fat pad aspirates. Therefore, ultrastructural evaluation of fat pad aspirates by electron microscopy should be utilized, given its increased sensitivity for amyloid detection. This article demonstrates a simple and reproducible procedure for performing anterior fat pad aspiration for the detection of amyloid utilizing both Congo red staining of cell block sections and electron microscopy for ultrastructural identification.

Fat pad aspiration is a preferred, minimally invasive, and low cost approach as compared to other methods to detect amyloid for diagnosis of systemic amyloidosis. This video article demonstrates a procedural outline for performing fat pad aspiration with appropriate processing of the specimen for the optimal diagnostic outcome.

表演和加工为淀粉样蛋白前脂肪垫活检

Historically, heart, liver, and kidney biopsies were performed to demonstrate amyloid deposits in amyloidosis.  Since the clinical presentation of this ...

科研

医学

Immunomagnetic Separation of Fat Depot-specific Sca1high Adipose-derived Stem Cells (ASCs)

The isolation of adipose-derived stem cells (ASCs) is an important method in the field of adipose tissue biology, adipogenesis, and extracellular matrix (ECM) remodeling. In vivo, ECM-rich environment consisting of fibrillar collagens provides a structural support to adipose tissues during the progression and regression of obesity. Physiological ECM remodeling mediated by matrix metalloproteinases (MMPs) plays a major role in regulating adipose tissue size and function1,2. The loss of physiological collagenolytic ECM remodeling may lead to excessive collagen accumulation (tissue fibrosis), macrophage infiltration, and ultimately, a loss of metabolic homeostasis including insulin resistance3,4. When a phenotypic change of the adipose tissue is observed in gene-targeted mouse models, isolating primary ASCs from fat depots for in vitro studies is an effective approach to define the role of the specific gene in regulating the function of ASCs. In the following, we define an immunomagnetic separation of Sca1high ASCs.

Immunomagnetic Separation of Fat Depot-specific Sca1high Adipose-derived Stem Cells ASCs

We present the techniques required to isolate the stromal vascular fraction (SVF) from mouse inguinal (subcutaneous) and perigonadal (visceral) adipose tissue depots to assess their gene expression and collagenolytic activity. This method includes the enrichment of Sca1high adipose-derived stem cells (ASCs) using immunomagnetic cell separation.

脂肪库专用SCA1的免疫磁性分离<sup&gt;高</sup&gt;脂肪干细胞（ASCs）

The isolation of adipose-derived stem cells (ASCs) is an important method in the field of adipose tissue biology, adipogenesis, and extracellular matrix ...

发育生物学

Human Skeletal Muscle Biopsy Procedures Using the Modified Bergstr&#246;m Technique

The percutaneous biopsy technique enables researchers and clinicians to collect skeletal muscle tissue samples. The technique is safe and highly effective. This video describes the percutaneous biopsy technique using a modified Bergstr&#246;m needle to obtain skeletal muscle tissue samples from the vastus lateralis of human subjects. The Bergstr&#246;m needle consists of an outer cannula with a small opening (&#8216;window&#8217;) at the side of the tip and an inner trocar with a cutting blade at the distal end. Under local anesthesia and aseptic conditions, the needle is advanced into the skeletal muscle through an incision in the skin, subcutaneous tissue, and fascia. Next, suction is applied to the inner trocar, the outer trocar is pulled back, skeletal muscle tissue is drawn into the window of the outer cannula by the suction, and the inner trocar is rapidly closed, thus cutting or clipping the skeletal muscle tissue sample. The needle is rotated 90&#176;&#160;and another cut is made. This process may be repeated three more times. This multiple cutting technique typically produces a sample of 100-200 mg or more in healthy subjects and can be done immediately before, during, and after a bout of exercise or other intervention. Following post-biopsy dressing of the incision site, subjects typically resume their activities of daily living right away and can fully participate in vigorous physical activity within 48-72 hr. Subjects should avoid heavy resistance exercise for 48 hr to reduce the risk of herniation of the muscle through the incision in the fascia.

The purpose of this video is to present the modified Bergstr&#246;m skeletal muscle biopsy technique on human subjects.

人骨骼肌活检手术使用修改伯格斯坦技术

The percutaneous biopsy technique enables researchers and clinicians to collect skeletal muscle tissue samples. The technique is safe and highly ...

Localization, Identification, and Excision of Murine Adipose Depots

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 reached a staggering sum of $190.2 billion in direct medical costs alone. Obesity is a major risk factor for a wide host of diseases. Historically, little was known regarding adipose and its major and essential functions in the body. Brown and white adipose are the two main types of adipose but current literature has identified a new type of fat called brite or beige adipose. Research has shown that adipose depots have specific metabolic profiles and certain depots allow for a propensity for obesity and other related disorders. The goal of this protocol is to provide researchers the capacity to identify and excise adipose depots that will allow for the analysis of different factorial effects on adipose; as well as the beneficial or detrimental role adipose plays in disease and overall health. Isolation and excision of adipose depots allows investigators to look at gross morphological changes as well as histological changes. The adipose isolated can also be used for molecular studies to evaluate transcriptional and translational change or for in vitro experimentation to discover targets of interest and mechanisms of action. This technique is superior to other published techniques due to the design allowing for isolation of multiple depots with simplicity and minimal contamination.

Due to the drastic and negative connection between obesity and other comorbidities, research on the role adipose plays in disease and overall health is warranted. We present a protocol for the isolation and excision of adipose depots allowing for the study of adipose using in situ and in vitro methods.

本地化识别，切除脂肪小鼠油库，并

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 ...

Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome

Percutaneous muscle biopsy using the Weil-Blakesley conchotome is well established in both clinical and research practice. It is a safe, effective and well tolerated technique. The Weil-Blakesley conchotome has a sharp biting tip with a 4 - 6 mm wide hollow. It is inserted through a 5 - 10 mm skin incision and can be maneuvered for controlled tissue penetration. The tip is opened and closed within the tissue and then rotated through 90 -180&#176; to cut the muscle. The amount of muscle obtained following repeated sampling can vary from 20 mg to 290 mg which can be processed for both histology and molecular studies. The wound needs to be kept dry and vigorous physical activity kept to a minimum for approximately 72 hr although normal levels of activity can restart immediately following the procedure. This procedure is safe and effective when close attention is paid to the selection of subjects, full asepsis and post procedure care. &#160;Both right and left vastus lateralis are suitable for biopsy dependent on participant preference.

Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome

This video demonstrates the technique of percutaneous muscle biopsy of the human vastus lateralis using the Weil-Blakesley conchotome.

人的<em&gt;股外侧肌</em&gt;骨骼肌活检使用韦尔 - Blakesley Conchotome

Percutaneous muscle biopsy using the Weil-Blakesley conchotome is well established in both clinical and research practice. It is a safe, effective and ...

Three-Dimensional Culture of Vascularized Thermogenic Adipose Tissue from Microvascular Fragments

Engineering thermogenic adipose tissue (e.g., beige or brown adipose tissues) has been investigated as a potential therapy for metabolic diseases or for the design of personalized microtissues for health screening and drug testing. Current strategies are often quite complex and fail to accurately fully depict the multicellular and functional properties of thermogenic adipose tissue. Microvascular fragments, small intact microvessels comprised of arteriole, venules, and capillaries isolated from adipose tissue, serve as a single autologous source of cells that enable vascularization and adipose tissue formation. This article describes methods for optimizing culture conditions to enable the generation of three-dimensional, vascularized, and functional thermogenic adipose tissues from microvascular fragments, including protocols for isolating microvascular fragments from adipose tissue and culture conditions. Additionally, best practices are discussed, as are techniques for characterizing the engineered tissues, and sample results from both rodent and human microvascular fragments are provided. This approach has the potential to be utilized for the understanding and development of treatments for obesity and metabolic disease.

Here, we present a detailed protocol outlining the use of microvascular fragments isolated from rodent or human fat tissue as a straightforward approach to engineer functional, vascularized beige adipose tissue.

微血管碎片血管化产热脂肪组织的三维培养

Engineering thermogenic adipose tissue (e.g., beige or brown adipose tissues) has been investigated as a potential therapy for metabolic diseases or for ...

生物工程

Clinical Protocol of Producing Adipose Tissue-Derived Stromal Vascular Fraction for Potential Cartilage Regeneration

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the knees by using different forms of mesenchymal stem cells (MSCs). In Korea, bone marrow concentrates and cord blood-derived stem cells have been approved by the Korean Food and Drug Administration (KFDA) for cartilage regeneration. In addition, an adipose tissue-derived stromal vascular fraction (SVF) has been allowed by the KFDA for joint injections in human patients. Autologous adipose tissue-derived SVF contains extracellular matrix (ECM) in addition to mesenchymal stem cells. ECM excretes various cytokines that, along with hyaluronic acid (HA) and platelet-rich plasma (PRP) activated by calcium chloride, may help MSCs to regenerate cartilage and improve knee functions. In this article, we presented a protocol to improve knee functions by regenerating cartilage-like tissue in human patients with OA. The result of the protocol was first reported in 2011 followed by a few additional publications. The protocol involves liposuction to obtain autologous lipoaspirates that are mixed with collagenase. This lipoaspirates-collagenase mixture is then cut and homogenized to remove large fibrous tissue that may clog up the needle during the injection. Afterwards, the mixture is incubated to obtain adipose tissue-derived SVF. The resulting adipose tissue-derived SVF, containing both adipose tissue-derived MSCs and remnants of ECM, is injected into knees of patients, combined with HA and calcium chloride activated PRP. Included are three cases of patients who were treated with our protocol resulting in improvement of knee pain, swelling, and range of motion along with MRI evidence of hyaline cartilage-like tissue.

Here, we present a protocol to produce an adipose tissue-derived stromal vascular fraction and its application to improve knee functions by regenerating cartilage-like tissue in human patients with osteoarthritis.

制备脂肪组织基质血管部分用于潜在软骨再生的临床协议

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the ...

Human Subcutaneous Adipose Tissue Sampling Using a Mini-Liposuction Technique

Studies on adipose tissue are useful in understanding metabolic and other conditions. Human subcutaneous adipose tissue is accessible. With appropriate training and strict adherence to aseptic technique, subcutaneous adipose samples can be safely and efficiently obtained in a non-clinical setting by researchers. Following the administration of local anesthetic lateral to the umbilicus, a 14 G needle attached to a 5 or 10 mL syringe is inserted through the skin into the subcutaneous tissue. Under suction, the syringe is moved in a reciprocating, slicing motion to isolate fragments of adipose tissue. Withdrawing the plunger is enough to ensure that adipose tissue fragments are aspirated through the needle into the syringe. A single biopsy can collect about 200 mg of tissue. This biopsy technique is very safe for both participants and research staff. Following the biopsy, participants can resume most everyday activities, although they should avoid swimming and overly strenuous activities for 48 h to avoid excessive bleeding. Participants can safely undergo 2 biopsies within a single day, meaning that the technique can be applied in before-after acute intervention studies.

The manuscript and associated video demonstrate a percutaneous biopsy technique to obtain samples of subcutaneous adipose tissue from areas surrounding the umbilicus. This method is a low-risk and efficient way to investigate a range of parameters (e.g., gene or protein expression, enzyme activity, lipid content) within adipose tissue.

使用微型吸脂技术进行人体皮下脂肪组织取样

Studies on adipose tissue are useful in understanding metabolic and other conditions. Human subcutaneous adipose tissue is accessible. With appropriate ...

Adipo-Clear: A Tissue Clearing Method for Three-Dimensional Imaging of Adipose Tissue

Adipose tissue plays a central role in energy homeostasis and thermoregulation. It is composed of different types of adipocytes, as well as adipocyte precursors, immune cells, fibroblasts, blood vessels, and nerve projections. Although the molecular control of cell type specification and how these cells interact have been increasingly delineated, a more comprehensive understanding of these adipose-resident cells can be achieved by visualizing their distribution and architecture throughout the whole tissue. Existing immunohistochemistry and immunofluorescence approaches to analyze adipose histology rely on thin paraffin-embedded sections. However, thin sections capture only a small portion of tissue; as a result, the conclusions can be biased by what portion of tissue is analyzed. We have therefore developed an adipose tissue clearing technique, Adipo-Clear, to permit comprehensive three-dimensional visualization of molecular and cellular patterns in whole adipose tissues. Adipo-Clear was adapted from iDISCO/iDISCO+, with specific modifications made to completely remove the lipid stored in the tissue while preserving native tissue morphology. In combination with light-sheet fluorescence microscopy, we demonstrate here the use of the Adipo-Clear method to obtain high-resolution volumetric images&#160;of an entire adipose tissue.

Due to the high lipid content, adipose tissue has been challenging to visualize using traditional histological methods. Adipo-Clear is a tissue clearing technique that allows robust labeling and high-resolution volumetric fluorescent imaging of adipose tissue. Here, we describe the methods for sample preparation, pretreatment, staining, clearing, and mounting for imaging.

Adipo: 三维脂肪组织成像的组织清除方法

Adipose tissue plays a central role in energy homeostasis and thermoregulation. It is composed of different types of adipocytes, as well as adipocyte ...

生物学

Human Adipose Tissue Micro-fragmentation for Cell Phenotyping and Secretome Characterization

In the past decade, adipose tissue transplants have been widely used in plastic surgery and orthopaedics to enhance tissue repletion and/or regeneration. Accordingly, techniques for harvesting and processing human adipose tissue have evolved in order to quickly and efficiently obtain large amounts of tissue. Among these, the closed system technology represents an innovative and easy-to-use system to harvest, process, and re-inject refined fat tissue in a short time and in the same intervention (intra-operatively). Adipose tissue is collected by liposuction, washed, emulsified, rinsed and minced mechanically into cell clusters of 0.3 to 0.8 mm. Autologous transplantation of mechanically fragmented adipose tissue has shown remarkable efficacy in different therapeutic indications such as aesthetic medicine and surgery, orthopedic and general surgery. Characterization of micro-fragmented adipose tissue revealed the presence of intact small vessels within the adipocyte clusters; hence, the perivascular niche is left unperturbed. These clusters are enriched in perivascular cells (i.e., mesenchymal stem cell (MSC) ancestors) and in vitro analysis showed an increased release of growth factors and cytokines involved in tissue repair and regeneration, compared to enzymatically derived MSCs. This suggests that the superior therapeutic potential of microfragmented adipose tissue is explained by a higher frequency of presumptive MSCs and enhanced secretory activity. Whether these added pericytes directly contribute to higher growth factor and cytokine production is not known. This clinically approved procedure allows the transplantation of presumptive MSCs without the need for expansion and/or enzymatic treatment, thus bypassing the requirements of GMP guidelines, and reducing the costs for cell-based therapies.

Here, we present human adipose tissue enzyme-free micro-fragmentation using a closed system device. This new method allows the obtainment of sub-millimeter clusters of adipose tissue suitable for in vivo transplantation, in vitro culture, and further cell isolation and characterization.

用于细胞表征和分泌特征的人类脂肪组织微分裂

In the past decade, adipose tissue transplants have been widely used in plastic surgery and orthopaedics to enhance tissue repletion and/or regeneration. ...