Abstract
Расширение панкреатических бета-клетках в естественных условиях или в силу естественных или поколения бета-клеток путем дифференцирования из эмбриона или взрослого стволовых клеток, может обеспечить новые расширяемые источники бета-клеток, чтобы облегчить дефицит доноров в трансплантации островков человека в качестве терапии для лечения диабета. Хотя недавние успехи были достигнуты к достижению этой цели, механизмы, регулирующие расширение бета и дифференцировку клеток из стволовых клеток / клеток-предшественников еще предстоит характеризуется. Здесь мы опишем протокол для модели повреждения в поджелудочной железе взрослых мышей, которые могут функционировать в качестве инструмента для изучения механизмов ремоделировании ткани и пролиферации бета-клеток и дифференциации. Частичное канал перевязки (PDL), является экспериментально индуцированной травмы поджелудочной железы грызунов участием хирургическое лигирование главного панкреатического протока в результате обструкции дренажа экзокринных продуктов из области хвоста поджелудочной железы. Нанесли ущерб вызывает атрофию ацинарных, иммунную infilt клетокРацион и тяжелой ткани ремоделирования. Ранее мы уже сообщали активации Neurogenin (NGN) 3, выражая эндогенных предшественников, как клетки и увеличение пролиферации бета-клеток после PDL. Таким образом, PDL обеспечивает основу для изучения сигналов, участвующих в динамике бета-клеток и свойства эндокринной прародителя во взрослой поджелудочной железы. Так, он по-прежнему остается в значительной степени неясным, какие факторы и пути способствуют регенерации бета-клеток и пролиферации в PDL, стандартизированный протокол для PDL позволит для сравнения между лабораториями.
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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