Musemodel af kirurgisk induceret Endometriose af Auto-transplantation af uterin væv
Summary
En beskrivelse af den kirurgiske induktion af endometriose hos mus og rotter ved auto-transplantation af uterine væv til arterielle kaskade af den intestinale mesenterium.
Abstract
Endometriose er en kronisk, smertefuld sygdom, hvis ætiologi er fortsat ukendt. Desuden kan behandlingen af endometriose kræver laparoskopisk fjernelse af læsioner, og / eller kroniske farmaceutiske behandling af smerter og infertilitet symptomer. Omkostningerne forbundet med endometriose er blevet anslået til 22 milliarder dollars om året i USA 1. For yderligere at vores forståelse af mekanismerne bag denne gådefulde sygdom, har dyremodeller været ansat. Primater spontant udvikler endometriose og derfor primater modeller nærmest ligner sygdommen hos kvinder. Gnaver modeller, er dog mere omkostningseffektive og let tilgængelige 2. Den model, som vi beskriver her, indebærer en autolog overførsel af uterin væv til tarmenes mesenterium (Figur 1) og blev første gang udviklet i rotter 3 og senere overført til musen 4. Målet med autologe gnaver model af kirurgisk induceret endometriose er at efterlignesygdommen hos kvinder. Vi og andre har tidligere vist, at den ændrede genekspression mønster observeret i endometriotic læsioner fra mus eller rotter, spejle, der blev observeret hos kvinder med sygdommen 5,6. En fordel ved at udføre operationen i musen er, at den overflod af transgene mus stammer ledigt. kan støtte forskere til at bestemme betydningen af specifikke komponenter vigtig i etablering og vækst af endometriose. En alternativ model, hvor fjernet menneskelige endometrie fragmenter bliver introduceret til bughinden af immunkompromitterede mus er også meget anvendt, men er begrænset af manglen på et normalt immunforsvar, som menes at være vigtigt i endometriose 2,7. Vigtigere er det, musemodel af kirurgisk induceret endometriose er en alsidig model, der er blevet brugt til at studere, hvordan immunsystemet 8, hormoner 9,10 og miljømæssige faktorer 11,12 påvirker endometriose samt virkningerne af endometriose på Fertility 13 og smerte 14.
Protocol
Discussion
Der er flere kritiske parametre, der bør bemærkes, mens de udfører de kirurgiske induktion af endometriose hos mus. Først endometriose er en østrogenafhængige sygdom, og som sådan denne operation skal udføres i intakte dyr eller alternativt i ovariectomized dyr suppleret med østrogener 20. For det andet skal suturering endometrie biopsier til arterielle kaskade udføres med største omhu. Vi har konstateret, at med kun to forholdsvis løs knob med én smide hver holder biopsi på plads, og samtidig f…
Disclosures
The authors have nothing to disclose.
Acknowledgements
Særlig tak til Chris Kassotis og Audrey Bailey til kritisk revision af dette manuskript og til Dr. Scott Korte, Joseph Beeman, Alison Curfman, Paul Kimball, Bridget Neibreggue, Jacob Redel, Amy Schroder, Maija Steinberg, og Stacey Winkeler for deres assistance i optimering af denne model i vores laboratorium. Finansieringen kom fra Klinisk Biodetectives Training Grant (NIH T90) (KEP), University of Missouri Life Sciences Undergraduate Research Muligheder Program, MU Forskningsråd, MU Research Board tilskud og NIH R21HD056441 (SCN).
Materials
| Name of the reagent/equipment | Company | Catalogue number |
| Wax pencil | Fisher | NC9954135 |
| Glass slide | Fisher | 12-550-433 |
| Eyedropper | Fisher | S79383 |
| Standard light microscope for evaluating vaginal cytology smears | ||
| Buprenorphine HCL c3 (CARJET) 10X1ml | Butler Animal Health Supply | 022891 |
| Sterile phosphate buffered saline (PBS) | Gibco | 14040-117 |
| 10,000U/ml Penicillin, 10,000μg/ml Streptomycin in 0.85% NaCl | Hyclone | SV30010 |
| Isoflurane | Abbott Animal Health | 05260-05 |
| Isoflurane non-rebreathing anesthetic system | ||
| Recirculating hot water heating pad | ||
| 30 ml syringe sheath | Fisher | 14-823-16G |
| Powder free sterile gloves | Fisherbrand | 19020558 |
| Ophthalmic ointment | Major Pharmaceuticals | 10033691 |
| Small electrical clippers | Wahl | 9861-600 |
| Chlorhexidine scrub | Fisher | NC9863042 |
| 70% Ethanol | ||
| Polylined sterile field | Busse Hospital Disposables | 696 |
| Size 3 scalpel | Fisher | 22-079-657 |
| Number 10 scalpel blades | Fisher | 22-079-681 |
| Small surgical scissors | Roboz | RS-5850 |
| Small serrated semi-curved forceps | Roboz | RS-5135 |
| 5-0 black braided silk suture | Ethicon | K870H |
| Sterilized pyrex glass Petri dishes | Corning | 70160-101 |
| 2 mm biopsy punch | Miltex | 33-31 |
| Sterile gauze | Kendall | 1806 |
| 6-0 black monofilament ethilon nylon suture | Ethicon | 697G |
| Needle drivers (optional) | World Precision Instruments | 500023 |
| 5-0 undyed braided coated vicryl suture | Ethicon | J490G |
| 9mm Autoclip wound clips | Becton Dickinson | 427631 |
| Autoclip applier & remover | Becton Dickinson | 427630 |
| 23G needle | Becton Dickinson | 305193 |
| 1cc syringe | Becton Dickson | 301025 |
| 5X magnifying glass stand (optional) | Fisher | 14-648-23 |
| 10% Buffered formalin | Fisher | SF100-4 |
| Calipers | Roboz | RS-6466 |
| Processing/embedding cassettes | Fisher | 15-197-700A |
| Biopsy foam pads | Fisher | 22-038-222 |
| RNAqueous RNA isolation kit | Ambion | AM1912 |
| Liquid nitrogen | ||
| Snap cap microcentrifuge flat top tube | Fisher | 02-681-240 |
| Ketamine (optional) | Simga | K4138 |
| Domitor (medetomidine hydrochloride) (optional) | Tocris | 2023 |
| Antisedan (atipamezole) (optional) | Sigma | A9611 |
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