Name: a 3d system for culturing human articular chondrocytes in synovial fluid
Uploaded: 2012-01-31T13:00:00
Description: Watch this Scientific Journal Video about A 3D System for Culturing Human Articular Chondrocytes in Synovial Fluid at JoVE.com

我々は、滑液の貯蔵及び遠心分離のヘルプを提供するためのロビンナイ（タフツ医療センター）、智也内村とダナケアンズ（タフツ大学）に感謝いたします。この作品は、LZのためのNIH（1R01AR059106 - 01A1）によって賄われていた

滑液の培養ヒト関節軟骨細胞用3Dシステム本研究では、我々は、修正された製造元で推奨されるカプセル化プロトコル（ロンザ、および31）を用いてアルギン酸ビーズで人間の関節軟骨細胞をカプセル化。これらの3次元構造体を使用して、我々は人間の滑液の様々な割合を含有する培養培地で細胞を培養するためのシステムを開発し、軟骨の遺伝子発現のための?...

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このレポートでは、我々は人間の滑液の高濃度を含む培地で、3D環境でのヒト関節軟骨細胞の培養を可能にする方法を開発した。滑液は関節軟骨細胞が存在する関節腔、自然環境を構成する主要コンポーネントの一つです。しかし、滑液の粘度は、軟骨細胞の三次元の長期培養するための大きな課題となっています。粘性環境での3D構造でさえ栄養分布を維持するための課題を克服し、凝集を...

特定の試薬と機器の表： <table border="1"><tr><th>試薬の名前</th><th>会社</th><th>カタログ番号</th><th>コメント</th></tr><tr><td>アルギン酸（アルギン酸ナトリウム塩） </td><td>シグマ</td><td> A2158 - 250G </td><td> 2.4％のソリューションは、40 ° Cで保存</td></tr><tr><td>粒状塩化カルシウム二水和物、 </td><td> JTベーカー</td><td> A19339 </td><td></td></tr><tr><td width="239" rowspan="2" >軟骨増殖培地</td><td width="254" rowspan="2" >ロンザ</td><td width="182" > CC - 3156（ベースのメ​​ディア） </td><td width="141" rowspan="2" ></td></tr><tr><td width="182" > CC - 4409（サプリメント） </td></tr><tr><td width="239" rowspan="2" >軟骨細胞分化のメディア</td><td width="254" rowspan="2" > Lonza </td><td width="182" > CC - 3226（ベースのメ​​ディア） </td><td width="141" rowspan="2" ></td></tr><tr><td width="182" > CC - 4408（サプリメント） </td></tr><tr><td>ヒト関節軟骨細胞</td><td>ロンザ</td><td > CC - 2550 </td><td></td></tr><tr><td> DAPI（4&#39;,6 -ジアミジノ-2 - フェニル二塩酸塩） </td><td>シグマアルドリッチ</td><td > D9542 </td><td></td></tr><tr><td> RNeasyミニキット（RNA抽出用） </td><td>キアゲン</td><td > 74104 </td><td></td></tr><tr><td> PCR試薬：SYBR -緑色</td><td>クアンタ</td><td > 95053-500 </td><td></td></tr><tr><td> 12mlの注射器</td><td>タイコ-ケンダルMonoject </td><td > 512852 </td><td></td></tr><tr><td> 22 - Gague注射針</td><td>タイコ-ケンダルL - Monoject </td><td > 8881 </td><td></td></tr><tr><td>顕微鏡</td><td>オリンポス</td><td > IX71 </td><td></td></tr><tr><td>プラットフォームのロッカー</td><td> Thermoscientific thermolyne </td><td >バリミックス</td><td></td></tr><tr><td></td><td></td><td ></td><td></td></tr><tr><td> プライマーの配列 </td><td></td><td ></td><td></td></tr><tr><td>コラーゲンIIaのフォワード</td><td> 5&#39; - TTC ATC CCA CCC TCT CAC AGT - 3&#39; </td><td ></td><td></td></tr><tr><td>コラーゲンIIA -逆</td><td> 5&#39; - CCTCTGCCTTGACCCGAA - 3&#39; </td><td ></td><td></td></tr><tr><td> MMP13フォワード</td><td> 5&#39; - TGT GCC CTT CTT CAC ACA GAC ACT - 3&#39; </td><td ></td><td></td></tr><tr><td> MMP13 -逆</td><td> 5&#39; - GAG AGC AGA CTT TGA GTC ATT GCC - 3&#39; </td><td ></td><td></td></tr><tr><td>カスパーゼ3フォワード</td><td> 5&#39; - TCA TTA TTC AGG CCT GCC GTG GTA - 3&#39; </td><td ></td><td></td></tr><tr><td>カスパーゼ3の逆</td><td> 5&#39; - TGG ATG AACのCAG GAG CCA TCC TTTの-3&#39; </td><td ></td><td></td></tr></table>

a 3d system for culturing human articular chondrocytes in synovial fluid

軟骨破壊は、変形性関節症の中心的な病理学的特徴、米国における障害の主要な原因です。成人の軟骨は、 生体内で非常に効率的に再生成されません、そしてその結果として、変形性関節症は軟骨の損失を不可逆的につながると慢性的な痛みと不動の1,2が付属しています。軟骨組織のエンジニアリングは、組織機能を再生成し、復元するために有望な可能性を提供しています。この技術は、通常、結果として得られる3Dは、エンジニアリングの目標生体 3-6 に欠損部位に移植することができる生化学的および生体力学的に成熟した組織で、ある期間にわたってバランスのとれた培地で構築する天然または合成の足場と培養に播種した軟骨細胞を含む。軟骨細胞の成長とマトリックスの沈着のための最適な条件を達成することが軟骨の組織工学の成功に不可欠です。 北極でのネイティブ関節腔、軟骨の骨のウラル表面は滑液に包まれている。この透明で粘性のある流体には、無血管性の関節軟骨に栄養を提供し、軟骨細胞の代謝7,8にとって重要な成長因子、サイトカインや酵素が含まれています。さらに、滑液は主に分泌する2つの主要なコンポーネント、ヒアルロン酸とlubricin 9 10を通じて、軟骨表面の間に低摩擦の移動が容易になります。対照的に、軟骨培養組織は、最も頻繁に人​​工培地で培養される。これらのメディアは、おそらく軟骨細胞代謝を研究するための複数の定義された条件を提供することができますが、関節液は、最も正確に関節軟骨細胞が入って置かれての自然環境を反映している確かに、滑液は、取得および保存するために容易であるという利点があり、多くの場合、定期的に体内で補充することができます。いくつかのグループが増殖するヒト、ウシ、ウサギおよびイヌcで滑液で培養液を補充しているhondrocytes、そのほとんどは滑液（20％以下）11月25日の唯一の低レベルを使用していました。鶏、馬と人間の軟骨細胞が滑液のより高い割合と培地で培養されているが、これらの培養システムは、2次元の26から28でした。ここでは、21日の期間にわたって滑液の割合が高く（最大100％）で、3Dシステムで培養ヒト関節軟骨細胞の我々の手法を提案する。そうすることで、我々は滑液の高粘度によって提示される主要なハードルを克服した。このシステムは、さらなるための自然環境を模倣するために軟骨のための自然環境を構成する2つの他の重要な要因（酸素張力および機械的荷重）29,30と組み合わせることができる3D設定で滑液中に人間の軟骨細胞を研究の可能性を、提供します。軟骨の成長。さらに、このシステムはまた、軟骨細胞の滑液の活性を測定するために使用し、開発するためのプラットフォームを提供することができます軟骨再生技術や関節炎の治療オプション。

Watch this Scientific Journal Video about A 3D System for Culturing Human Articular Chondrocytes in Synovial Fluid at JoVE.com

A 3D System for Culturing Human Articular Chondrocytes in Synovial Fluid

滑液の高濃度の培養ヒト関節軟骨細胞の3次元システムについて説明する。滑液は関節軟骨のための最も自然な微小環境を反映し、容易に入手し、保存することができます。このシステムは、このように軟骨再生の研究や、関節炎を治療するための治療薬をスクリーニングするために使用することができます。

滑液における培養ヒト関節軟骨細胞用の3Dシステム

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