Abstract
胚または成体幹細胞からの分化によってin vivoまたはex vivoで 、またはβ細胞の発生における膵臓β細胞の拡大は、糖尿病の治療法として、ヒト膵島 移植におけるドナーの不足を緩和するβ細胞の新しい拡張可能な供給源を提供することができます。最近の進歩は、この目標に向けてなされてきたが、幹細胞/前駆細胞からβ細胞の増殖および分化を制御する機構を特徴づけされていません。ここでは、組織リモデリングおよびβ細胞の増殖および分化のメカニズムを研究するためのツールとして機能することができる成体マウスの膵臓における傷害モデルのためのプロトコルを説明します。部分管結紮（PDL）は、膵臓の尾部領域のうち外分泌物の排水の閉塞をもたらす主膵管の外科的結紮を伴うげっ歯類膵臓の実験的に誘導損傷です。被害は腺房萎縮、免疫細胞infiltを誘導します配給、重度組織再構築。我々は以前、内因性前駆細胞様細胞およびPDL後のβ細胞増殖の増加を表現するニューロゲニン（NGN）3の活性化を報告しています。したがって、PDLは、ベータ細胞の動態に関与するシグナルと成人膵臓内分泌前駆細胞の性質を研究するための基礎を提供します。 、それはまだ要因と経路がPDLにおけるβ細胞新生および増殖に寄与する、大部分は不明のままであるので、PDLのための標準化されたプロトコルは、研究室全体の比較を可能にします。
Materials
| Name | Company | Catalog Number | Comments |
| Supplies for preparation area | |||
| Hibiscrub | Regent Medical | 5601IE5F11 | Chlorhexidin diglucon |
| Ketamin | Ceva | BE-V202526 | anesthesia |
| Rompun(2%) (Xylazin) | Bayer | BE-V041815 | anesthesia |
| Duratears | Alcon | 34335-8 | ointment for eyes |
| Razor | |||
| Supplies for surgical area | |||
| Leica Operating microscope | Leica | 10446320 | |
| Hot bead sterilizer | Fine Science Tools (FST) | 18000-45 | |
| autoclaved surgical instruments | Fine Science Tools (FST) | ||
| Recirclulating water heating pad | Gaymar Industries, Inc. | TP702 | |
| Adhesive OP-towel | BARRIER | 706500-07 | |
| OP-tape | BARRIER | 381035-00 | |
| Stella 3/5 Compresse de gaze (Sterile) | Lohmann&Rauscher International | 35968 | |
| Mini Plasco 0.9% NaCl solution | B.BRAUN | 3521680 | |
| 6-0 prolene suture | Ethicon | 8706H | |
| 4-0 polyglycol suture | Ethicon | SL-607 | |
| Supplies for recovery area | |||
| Paper bedding (paper towel) | |||
| Vetergesic | ecuPhar | BE-V342955 | |
| Materials for analysis | |||
| Taqman Ngn3 primer | Integrated DNA technologies | Mm.PT.56a.33574796.gs | |
| Taqman CycloA primer | Integrated DNA technologies | Mm.PT.39a.2.gs | |
| anti-Rabbit Cleaved Caspase 3 antibody (D175) | Cell Signaling | 5A1E | |
| anti-mouse/rat Ki-67 antibody | eBioscience | 14-5698-82 | |
| monoclonal anti-actin alpha-Smooth muscle-Cy3 (mouse) | Sigma | 085K4889 | |
| anti-rat Cytokeratin 19 | DSHB | ||
| monoclonal Mouse anti-BrdU | Dako | M0744 | |
| Hoechst | Sigma | 33342 |
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