포털 정맥 주입: 간암 전이를 연구하는 방법

Abstract

출처: 고다드, E. T., 외. 유방암의 간 전이를 연구하는 포털 정맥 주입 모델. J. Vis. Exp. (2016).

이 비디오는 포털 정맥 주사를 통해 뮤린 간으로 유방 종양 세포를 전달하는 외과 적 절차를 설명합니다. 이 모델은 간 전이의 늦은 단계의 조사를 허용합니다.

Protocol

1. 수술 부위 및 기기 준비 계획된 수술 전에 124°C에서 30분, 1 -2일 동안 124°C에서 자동으로 자동화하여 가위, 집게 및 헤모스타를 준비한다. 수술 후 회복을 위해 자동 식용 또는 멸균 침구, 케이지 및 음식에 대한 액세스를 보장하십시오. 기동 후드에서 기부 수술 부위를 준비한다. 가열 패드, 광원, 마취 관 및 코 콘을 포함하여 10 % 표백제로 수술 영역의 모든 표면…

Materials

1 ml Syringe w/ 26-gauge Needle	BD Syringe	309597	309597
Alcohol Prep Pads	Fisher Scientific	06-669-62	For cleaning of abdomen prior to surgical incision
All Purpose Sponges, Sterile	Kendall	8044	4" x 4", use dipped in sterile saline to keep large and small intestines protected and hydrated during surgery
Artificial Tears	Rugby	370114	Mineral oil 15%, white petrolatum 83%; use to protect eyes during surgery
Buprenorphine HCl, 0.3 mg/ml	Mfg. by Reckitt Benckiser	NDC-12496-0757-1	Use at 0.05 – 0.1 mg/kg body weight, 1 – 2x daily for 72 hr, injected subcutaneously
Bupivacaine HCl, 0.5% (5 mg/ml)	Mfg. by Humira Inc	NDC-04091163-01	Use at 0.5%, 1x immediately after surgery, 10 μl injected subcutaneously at incision site
Celox™ Rapid Hemostatic Gauze	Medtrade Products Ltd.	FG08839011	Cut into 5 mm² pieces, use to stop blood flow out of the portal vein with pressure following injection
Chlorhexidine, 2% Solution	Vet One	1CHL008	Use caution, do not get chlorhexidine in mucous membranes or ears of the mouse
Cotton Tipped Applicators, Sterile	Fisher Scientific	23-400-114	6" Wooden Shaft 2 pc/envelope
DMEM, High-Glucose	HyClone	SH30243.01	Cell culture media base for use with D2A1, D2.OR, and 4T1 mammary tumor cell lines
Dry Glass Bead Sterilizer			Use between surgeries to sterilize stainless steel tools, use caution, extremely hot; multiple suppliers
Ethanol, 70% solution			Use caution flammable; use to clean surgical area as needed; multiple suppliers
Fetal Bovine Serum	HyClone	SH30071.03	Cell culture media additive for use with D2A1, D2.OR, and 4T1 Journal of Visualized Experiments www.jove.com Copyright © 2016 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License Page 2 of 3 mammary tumor cell lines, use at 10% in DMEM high glucose
Gauze, Sterile	Kendall	2146	2" x 2", use dipped in chlorhexidine 2% solution for cleaning of abdomen prior to surgical incision
Isoflurane	Piramal	NDC-66794-017-25	Administered at 2.5%
Isoflurane Vaporizer	VetEquip	911103	Use caution, vaporizes anesthetic gases
Removable Needle Syringe, 25 μl, Model 1702	Hamilton	7654-01	For portal vein injection; use caution, paricularly while working with tumor cell-loaded needles, sharp when needle is attached
Scalpel handle			Stainless steel; multiple suppliers
Scalpel blade, #15			Carbon steel, sterile, size 15; multiple suppliers
Small Hub Removable Needles, 32-gauge	Hamilton	7803-04	For portal vein injection, 1" length, point style 4, 12° angle, 33- to 34- gauge reusable needles can also be used; use caution, paricularly while working with tumor cell loaded needles, sharp
Sterile Saline	Fisher Scientific	BP358-212	0.9% NaCl solution; alternatively, can be homemade and sterile filtered
Surgical Gloves, Sterile			Multiple suppliers
Sutures, Sterile	Ethicon	J310H	4-0 27" coated vicryl w/ 22 mm 1/2c taper ethalloy needle; use caution, sharp
Table Top Portable Anesthesia Machine	VetEquip	901801	Use with isoflurane vaporizer for mouse anesthesia
Thumb Dressing Forceps			Stainless steel, serrated, blunted; multiple suppliers
Towel Drapes, Sterile	Dynarex	4410	18" x 26", to cover heating pad and provide a sterile workspace during surgery