この作品は、欧州連合のホライゾン 2020 (グラント契約号 645710) と第 7 フレームワークから (グラント契約番号 612547 と 319010) のプログラムの資金を受信しました。著者生検の手順と支援の博士フォティーニ Sourli Chasioti を感謝したいです。著者らはまた、ビデオの準備で彼の貴重な助け氏 Yiorgos Paterakis を感謝します。

研究ヘルシンキ宣言によって設定された基準に適合し、運動科学部テッサリア大学 Βiοethics 委員会によって承認されました。
 1 です手術機器や消耗品の準備 <ol><li>4輪手術メイヨーのスタンドに消毒手術フィールドの位置。</li>。
	<li>4輪手術メイヨーのスタンドに以下の消耗品・手術用機器の位置: 
	ストレート営業はさみ (15 cm) の 1 つのペア 1 メス (11 号) 1 メス ハンドル 曲線はさみ (14 cm) の 1 つのペア 1 つモスキート鉗子 コッヘル ピンセット 1 組 1 つの外科ピンセット ポビドン ヨード (の生殖ガーゼ) One 縫合 (4.0) ストレートはさみ (11 cm) の 1 つのペア 1 つ針ホルダー (15 cm) 5 滅菌ガーゼ 1 つの接着剤滅菌ガーゼ 消毒された針を用いて 1 つの 10 mL シリンジ 2% キシロカイン (ないアドレナリン) 10 mL 1 つ消毒手術用手袋のペア</li> <li>別のベンチに以下の材料を位置: 2 つのマイクロ遠心チューブ用 10% ホルマリン 5 mL を含む 1 つの管 50-100 mL の-190 ° C で液体窒素を含んでいる 1 つの小さい容器</li></ol> 2。参加者の準備 <ol><li>参加者 15 生検の手順の前に 8 時間高速に従います</li>。
	<li>参加者を控える運動、過度のストレス、アルコール、およびアクティブおよび受動喫煙 72 h 前生検 16 , 17 を持っている</li>。
</ol> 3。皮下脂肪組織の生検を行う <ol><li>仰臥位で手術のベッドの上、参加者を配置します</li>。
	<li>ポビドン ヨードを使用して切開部を消毒します</li>。
	<li>ゆっくりと着実に 2% キシロカイン (ないアドレナリン) 10 mL を注入局所麻酔として使用する選択した領域に 18.</li>
	<li>切開部を除く参加者の体をカバーする領域上で滅菌手術フィールドを配置します</li>。
	<li>少なくとも 3-5 分は、その後、ピンセットのペアを持つ局所麻酔の領域に圧力を適用、彼の感じているその局所麻酔が達成されていることを確認する任意の痛み 19 参加者を求める</li>。
	<li>号 11 メスとメス ハンドルを付ける</li>。
	<li>へその右または長さ 2 〜 2.5 cm、左に 3-5 cm の切開を作成します</li>。
	<li>営業はさみストレート 15 cm のペアで皮下組織を削除します</li>。
	<li>脂肪組織を明らかにすると、一度はピンセットのペアと湾曲した 14 cm はさみのペアでそれをキャプチャします</li>。
	<li>カット約 1 g の脂肪組織を削除します</li>。
	<li>次の組織の除去、止血して切開に滅菌ガーゼを置きます</li>。
	<li>止血を確認すると、キャプチャ 14 cm 針で 4.0 の縫合と皮膚外科ピンセットのペアと皮下組織が表示されるまでします</li>。
	<li>皮膚や皮下に密接に並べながら皮下組織へ移動する、皮膚に縫合針を挿入します</li>。
	<li>縫合針、皮下組織、皮膚を外部から駆動するまでに連続ジグザグ法に従います。皮膚切開が閉じられるまで継続します</li>。
	<li>外部から縫合糸を結ぶ</li>。
	<li>生理食塩水で切開部をきれいにし、接着剤の滅菌ガーゼでカバーします</li>。
</ol> 4。組織の沈着 <ol><li>3枚に収集した脂肪組織を分ける: 150、350 ω、500 mg</li>。
	<li>マイクロ遠心チューブ用のサンプル組織の 150 および 350 mg の作品を配置します</li>。
	<li>-80 で冷凍庫の最終的な沈着まで-190 ° C で液体窒素で小型の容器にマイクロ遠心チューブを用ひたる 2 ° C 
	注: 事前に-190 で液体窒素中でマイクロ遠心チューブ用の浸漬中カップでの失敗を避けるためにマイクロ遠心チューブ用の小さな穴をパンクに滅菌針を使用 ° C</li>
	<li>500 mg のサンプル組織片を浸し-組織学的解析に使用する-10% ホルマリン 5 mL を含む管内 。
	注: は、手術中に出血した場合にのみモスキート鉗子を使用します。この場合において、結紮して止血が達成されるまでを閉じるモスキート鉗子で船をキャプチャします</li>。
</ol>

<ol>
	<li>Bruening, W., 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=Systematic+review:+comparative+effectiveness+of+core-needle+and+open+surgical+biopsy+to+diagnose+breast+lesions.">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="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+sampling+and+long-term+storage+of+subcutaneous+adipose-tissue+biopsies+for+determination+of+fatty+acid+composition.">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://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+and+adipose+tissue+biopsy+in+older+adults+with+type+2+diabetes+.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Needle+and+surgical+biopsy+techniques+differentially+affect+adipose+tissue+gene+expression+profiles.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnostic+value+of+abdominal+wall+fat+pad+biopsy+in+senile+systemic+amyloidosis.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+biopsy+method+to+increase+yield+of+subcutaneous+abdominal+adipose+tissue.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+wedge+and+needle+hepatic+biopsy+in+open+bariatric+surgery.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+surgical+treatment+of+the+metabolic+syndrome+on+liver+fibrosis+and+cirrhosis.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+single-blind+controlled+study+of+electrocautery+and+ultrasonic+scalpel+smoke+plumes+in+laparoscopic+surgery.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+Diathermy.">Surgical Diathermy.</a> Br J Hosp Med. 52, (1994).</li><li>Tomita, 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=Mutagenecity+of+smoke+condensates+induced+by+CO-laser+irradiation+and+electrocauterization.">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="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterisation+and+removal+of+electrosurgical+smoke.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ligasure+versus+diathermy+scissors+tonsillectomy:+A+controlled+randomized+study.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemostasis+in+thyroid+surgery:+harmonic+scalpel+versus+other+techniques--a+meta-analysis.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+pilot+study+of+sampling+subcutaneous+adipose+tissue+to+examine+biomarkers+of+cancer+risk.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+a+single+bout+of+exercise+on+resting+energy+expenditure+and+respiratory+exchange+ratio.">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="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intra-individual+variation+of+basal+metabolic+rate+and+the+in&#64258;uence+of+daily+habitual+physical+activity+before+testing.">Intra-individual variation of basal metabolic rate and the in&#64258;uence 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.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infiltrative+anesthesia+in+office+practice.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasma+levels+of+2%+lidocaine+with+1:100,000+epinephrine+with+young+children+undergoing+dental+procedures.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+Aging+on+Inflammation+and+Hemostasis+through+the+Continuum+of+Critical+Illness.">Effects of Aging on Inflammation and Hemostasis through the Continuum of Critical Illness.</a> Aging Dis. 2, 501-511 (2011).</li></ol>

著者は、彼らは競合する金銭的な利益があることを宣言します。

本研究の目的は、健康な男性の非ジアテルミー法を用いた局所麻酔下で実行、腹部皮下脂肪の生検のための手術手技の標準化をだった。ジアテルミー、オペレーティング ルームで使用されます。ただし、可能性のある副作用 (すなわち、フラッシュ火災と環境毒性)9と10の治癒の遅れが発生します。ジアテルミーのこれらの不必要な効果は、営業職員...

脂肪組織生検は、代謝と疾患に関するメカニズムの洞察力に富んだ概要を提供可能性があります分析、組織サンプルを提供しています。例えば、乳房癌腫瘍同定1、脂肪酸組成試験2、重要な代謝とホルモンの病気経路3の研究に使用されます。腹部の皮下脂肪組織生検を得るためには、研究者や医師は、手術または針ベースの手法を使用します。しかし、手術皮下脂肪の生検は、複雑な生物学的メカニズム[メッセンジャーのリボ核酸 (mRNA) 式など、組織学的解析等]のより包括的な概要を提供することが組織サンプルを提供できます4.彼らは健康な成人4で使用されている以前とベッドサイド5で安全に実行できます。
針による生検法は組織の整合性を破壊する強烈な脂肪吸引を必要とし、それは通常組織 (100-500 mg)6の少量を取得します。これら組織サンプルは、複雑な研究のために適切かもしれない。さらに、針ベースの手法と比較して、手術皮下脂肪の生検を提供できる (例えば、 B および T 細胞シグナリング); いくつかの炎症性指標のより完全な絵を描くことがあります組織サンプル細胞骨格の調節;携帯電話経路;脂質、炭水化物およびアミノ酸の代謝経路酸化的リン酸化経路4。また、針によるには組織4,報7,8 の線維化領域について少し情報を提供しながらの手術 1 つ4より血液細胞汚染のリスクが高いが含まれて.
通常、手術の脂肪組織生検には9過剰な出血を防ぐために血管を焼灼のジアテルミー治療が含まれます。ジアテルミーは、燃やすし、腫瘍や疣贅病9で感染している組織によって影響を受ける組織の破壊にも使用されます。この手法は、特に脳神経や目の手術9に該当します。
ジアテルミーの過度の使用は、壊死組織、感染の原因と治癒過程10を遅らせることの形成につながる可能性があります。確かに、次のジアテルミー9フラッシュ火災など皮膚病変組織の副作用を報告されています。さらに、参加者/患者と営業スタッフ9によって吸入されることがあります手術の環境放出のジアテルミー手術煙の毒性について懸念を提起されています。確かに、ジアテルミーの使用、喫煙 6 タバコ11によって生成された同じ手術室空気毒性を引き起こす可能性があり、少なくとも 20 分12 汚染することができます最新の手術室は通常、十分な換気システムを持っていますが、.しかし、扁桃手術患者、ジアテルミー法少ないが、有意ではない、術後の痛み13と古典的な結紮止血法より出血14は示します。全体的にみて、手術後の痛みの違い、かろうじて健常者に脂肪組織生検後の率の出血の証拠が存在します。証拠の欠如とジアテルミーの副作用を考えると、脂肪組織の生検のため非ジアテルミー法の標準化が必要です。したがって、本研究の目的は健康な男性で非ジアテルミー法を用いた局所麻酔下で腹部の皮下の脂肪組織生検のための手術手技の標準化をだった。さらに、我々 は組織コレクション mRNA の発現および蛋白質の集中を決定し、組織学的解析を実行するための手順を概説します。

<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

脂肪組織生検は、代謝と疾患に関するメカニズムの洞察力の概要を提供することが分析、組織サンプルを提供しています。腹部の皮下脂肪組織生検を得るためには、研究者や医師は、手術または針ベースの手法を使用します。しかし、手術皮下脂肪の生検は白色脂肪組織の生物学的指標の複雑さのより包括的な概観を提供することが組織サンプルを提供できます。通常、手術の脂肪組織生検には、過度の出血を防ぐために血管を焼灼ジアテルミー治療が含まれています。それにもかかわらず、ジアテルミー後フラッシュ火災、体内、皮膚病変などの副作用が報告されています。したがって、非ジアテルミー法を用いた局所麻酔下で腹部の脂肪組織を外科的生検の標準化を目的とします。非ジアテルミー腹部外科的生検法による健康な男性で 115 の皮下脂肪の生検を行った.我々 の結果は、115 事業 (2.61%) から出血過剰なポスト処理の 3 つのケースを示した。結論としては、最小限の副作用で、ベッドサイドでの健康的な男性に非ジアテルミー メソッドを使用して、標準化された腹部皮下脂肪外科的生検を安全に適用できます。

参加者の特性は、表 1に提供されます。切開部の縫合は、8-12 日以内から落ちた。我々 は健常成人男性で非ジアテルミー腹部外科的生検法 115 の皮下脂肪組織生検の合計を完了した (年齢: 35.2 ± 6.9 年; ボディマス指数: 27.3 ± 4.4; および体脂肪率: 27.0 ± 9.0%)。3 つのケースで観察された後生検出血 (有病率: 2.61%)、手術後 2 日間。過度の出血を経験した 3 人は?...

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

我々 は局所麻酔下で非ジアテルミー法を用いた腹部脂肪組織生検を標準化しました。115 事業 (2.61%) から出血過剰な術後の症例が発生しました。健康な男性に非ジアテルミー法を用いた腹部脂肪組織の外科的生検を安全に適用できることが分かった.

非ジアテルミー法による腹部皮下脂肪の生検のためのテクニック

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

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

Research

JoVE Journal

Medicine

15.0K ビュー.  University of Thessaly. 我々 は局所麻酔下で非ジアテルミー法を用いた腹部脂肪組織生検を標準化しました。115 事業 (2.61%) から出血過剰な術後の症例が発生しました。健康な男性に非ジアテルミー法を用いた腹部脂肪組織の外科的生検を安全に適用できることが分かった.

ビデオ: 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.

アミロイドのための前方脂肪パッドのFNAの実行と処理

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

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.

脂肪デポ固有のSca1の免疫磁気分離<sup&gt;高</sup&gt;脂肪由来幹細胞（ASCに）

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

発生生物学

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.

変更されたベルイストローム法を用いたヒト骨格筋生検手順

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.

マウス脂肪デポのローカライズ、識別、および切除

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.

人間<em&gt;外側広筋</emワイル・ブレークスリーConchotomeを使用&gt;骨格筋生検

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.

微小血管断片からの血管新生熱発生脂肪組織の三次元培養

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

生物工学

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.

潜在的な軟骨再生のための脂肪組織由来間質血管分数の生産の臨床プロトコル

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.

ミニ脂肪吸引法を用いたヒト皮下脂肪組織サンプリング

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.

裏打ちクリア: 組織の脂肪組織の三次元イメージング法をクリア

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.

細胞表現型と分泌性特性化のためのヒト脂肪組織微細断片化

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