Name: isolation and culture of dental epithelial stem cells from the adult mouse incisor
Uploaded: 2014-05-01T13:00:00
Description: Watch this Scientific Journal Video about Isolation and Culture of Dental Epithelial Stem Cells from the Adult Mouse Incisor at JoVE.com

著者は、DESCの文化と初期支援する秀のping王に感謝します。著者は（ODKにK12-GM081266は、MGCに、K99-DE022059 AHJにK08-DE022377-02 OHに、そしてR01-DE021420）国立衛生研究所からの奨学金や助成金によって部分的に資金を供給している。

1。成体マウスから低ヘミ顎を分析<ol><li>このプロトコルを実行する前に、必要な制度の承認を得て、すべての動物保護ガイドラインに準拠するようにしてください。標準動物実験委員会の手続きを承認用いて動物を安楽死させる。この作業のために、CO 2窒息頸椎脱臼に続いて使用されました。 <ol><li>オプション：カミソリの刃を使用して、身体の残りの部分とは別のヘッド。 </li><...

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上皮細胞は、40年以上前に21〜24第一に成功培養し、さらに最近では、上皮幹細胞の正常な分離25〜27は、上皮生物学の知識を進んでいる。我々は、成体マウスの切歯のDESCs、歯の生物学とエナメル質の形成に重要な洞察をもたらす可能性を秘めている幹細胞の比較的代役集団を単離するためのプロトコルを報告している。このプロトコルは、最初は毛包28から上皮幹細?...

脊椎動物のユニークな特徴の一つは、歯の進化である。この器官に関連する分子経路及び形態学の専門分野が発達し、進化生物学者5によるものを含む、いくつかの観点から検討されているように歯は、研究の多くの分野のための重要なモデルシステムとなっています。より最近では、再生医療の分野は、モデルとしての歯を使用して貴重な洞察を得るために始めている。特に、歯科用上皮幹細胞の発見は、重要な進歩であった6-13。 すべてのげっ歯類は、その成長のこれらの歯の成体幹細胞制御を研究するためのアクセス可能なモデルシステムを作り、幹細胞によって支えられて継続的に成長している切歯を有する。遺伝系統トレース実験8,9,12,14が続き、1970年代10,11における標識実験は、DESCsは切歯の近位領域に存在することを明らかにした。ステム細胞唇子宮頸ループ（CL）として知られている推定のニッチの外唇側に移動、上の上皮の区画に子孫、およびトランジット増幅（TA）細胞（ 図1）と呼ばれる細胞の集団に貢献しています。具体的には、DESCsは、外エナメル上皮（OEE）に存在し、星状胞体（SR）8,9,14、および内エナメル上皮（IEE）は、細胞周期の限られた数を経て進行してから移動し、TA細胞を生じさせる遠位側切歯の長さ（ 図1）に沿って。マウス切歯で分化エナメル芽細胞は、1日15,16で約350ミクロンの驚くべき割合で切歯に沿って遠位方向に移動し続けています。彼らが移動すると、細胞が成熟したエナメル芽細胞と角質アラト（SI）の細胞に分化する。エナメルマトリックスの全層を堆積した後、エナメル芽細胞の多くは、アポトーシスを受け、残りのセルのサイズが縮小し、エナメル質の成熟を制御する&lt;&gt; 17 SUP。このようなSr等の唇のCLでの他の細胞型の系統は、それほど明確であり、間充織18内および舌のCLでの幹細胞に関するデータのみ出現し始めている。 マウス切歯モデルを使用して、グループの数は、遺伝的経路と自然の幹細胞に基づいた臓器再生に関与する細胞生物学的プロセスを解明するために取り組んできました。しかし、唇CLが困難な初代細胞での作業になり約5000のマウス切歯あたりと推定された細胞の数が比較的少なく、含まれています。そのため、努力が文化に作られ、 生体内で達成可能ではない実験的なアプローチに新たな扉を開くために体外 6,16,19,20 におけるこれらの細胞を拡大してきた。ほとんどの最近の研究は、これらの細胞が両方の自己再生および培養13発現細胞アメロゲニンに分化できることを実証した。ここでは、目のための方法を説明し、実証するマウス唇CLからの細胞のE信頼性と一貫性の文化。

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			<td height="63" style="height:63px;width:216px;">Forceps, size 5</td>
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isolation and culture of dental epithelial stem cells from the adult mouse incisor

歯の再生と刷新の根底にある細胞および分子メカニズムを理解することは、大きな関心1-4の話題になると、マウスの切歯は、これらのプロセスのためのモデルを提供しています。この驚くべき器官は、動物の生涯を通じて継続的に成長し、（舌側）（唇側）唇と舌頚椎ループ（CL）領域内に収容された成体幹細胞のアクティブなプールから必要なすべての細胞型を生成します。唇CLからのみの歯科幹細胞は、唇表面のエナメル質、歯の外側カバーを、生成エナメル芽細胞を生じさせる。この非対称エナメル形成は切歯先端に擦り傷を可能にし、近位の切歯における前駆細胞や幹細胞は歯の組織が常に補充されていることを確認。 試験管内で 、これらの前駆細胞もしくは幹細胞を単離および増殖する能力は、その拡張を可能にし、例えばHなどの生体内では達成できない多くの実験、への扉を開きます潜在的な幹細胞調節因子のIGHスループット·テスト。ここでは記述し、マウスの切歯の唇CLからの培養細胞への信頼性と一貫性のある方法を示す。

マウスヘミ-下顎骨の1つを連続的に成長切歯三大臼歯根付いた（ 図1A）を含む。すべての歯は、象牙質とエナメル質、歯冠の2鉱化コンポーネント（ 図1Aおよび1A &#39;）で構成されています。切歯を収容2つのステム細胞ニッチは、唇および舌CL CLと呼ばれ、エナメル質は、唇側（ 図1A &#39;）のみに形成されている。 DESCsは切歯エナメル質の形成...

Watch this Scientific Journal Video about Isolation and Culture of Dental Epithelial Stem Cells from the Adult Mouse Incisor at JoVE.com

Isolation and Culture of Dental Epithelial Stem Cells from the Adult Mouse Incisor

成体マウス切歯から歯科上皮幹細胞の単離および培養

継続的に成長しているマウスの門歯は、歯科上皮幹細胞（DESCs）から歯の組織の更新を研究するためのモデルを提供します。一貫して確実切歯からこれらの細胞を取得し、in vitroでそれらを拡張するための堅牢なシステムをここで報告されている。

Understanding the cellular and molecular mechanisms that underlie tooth regeneration and renewal has become a topic of great interest1-4, and the mouse ...

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Biology

Adult and Embryonic Skeletal Muscle Microexplant Culture and Isolation of Skeletal Muscle Stem Cells

大人と胚骨格筋のMicroexplantの文化と骨格筋幹細胞の単離

Cultured embryonic and adult skeletal muscle cells have a number of different uses. The micro-dissected explants technique described in this chapter is a ...

Isolation of Retinal Stem Cells from the Mouse Eye

マウスの眼から網膜幹細胞の単離

The adult mouse retinal stem cell (RSC) is a rare quiescent cell found within the ciliary epithelium (CE) of the mammalian eye1,2,3. The CE is made up of ...

Isolation and Culture of Adult Epithelial Stem Cells from Human Skin

ヒト皮膚から大人上皮幹細胞の単離と培養

The homeostasis of all self-renewing tissues is dependent on adult stem cells. As undifferentiated stem cells undergo asymmetric divisions, they generate ...

Generation of Mice Derived from Induced Pluripotent Stem Cells

人工多能性幹細胞に由来するマウスの作製

The production of induced pluripotent stem cells (iPSCs) from somatic cells provides a means to create valuable tools for basic research and may also ...

Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle

成人骨格筋からの個々の筋線維とその衛星細胞の単離および培養

Muscle  regeneration in the adult is performed by resident stem cells called satellite  cells. Satellite cells are defined by  their position between the ...

Na&iuml;ve Adult Stem Cells Isolation from Primary Human Fibroblast Cultures

NaÃƒÆ’Ã‚Â¯ve Adult Stem Cells Isolation from Primary Human Fibroblast Cultures

初代ヒト線維芽細胞培養からナイーブ成人幹細胞の分離

Over the last decade, several adult stem cell  populations have been identified in human skin 1-4.  The isolation of multipotent adult dermal precursors ...

Induction and Analysis of Epithelial to Mesenchymal Transition

間葉転換を誘導し、上皮の分析

Epithelial  to mesenchymal transition (EMT) is essential for proper morphogenesis during  development. Misregulation of this  process has been implicated ...

Isolation, Culture and Transduction of Adult Mouse Cardiomyocytes

成体マウスの心筋細胞の単離、培養および形質導入

Cultured cardiomyocytes can be used to study cardiomyocyte biology using techniques that are complementary to in vivo systems. For example, the purity and ...

Isolation and Culture of Primary Mouse Keratinocytes from Neonatal and Adult Mouse Skin

新生児および成人マウス皮膚由来の初代マウスケラチノサイトの単離および培養

The keratinocyte (KC) is the predominant cell type in the epidermis, the outermost layer of the skin. Epidermal KCs play a critical role in providing skin ...

Isolation of Enteric Glial Cells from the Submucosa and Lamina Propria of the Adult Mouse

粘膜と粘膜固有層において大人のマウスから腸のグリア細胞の分離

The enteric nervous system (ENS) consists of neurons and enteric glial cells (EGCs) that reside within the smooth muscle wall, submucosa and lamina ...