Isolated limb perfusion with melphalan for human melanoma xenografts in the hindlimb of nude ratscolon; a surviving animal model

摘要

We have established a surviving model of isolated limb perfusion using xenografts of the human melanoma cell line MM 96L injected subcutaneously into the hindlimb of a nude rat. The femoral artery and vein were cannulated via the left renal artery and vein and the hind limb was isolated using tourniquets. The limb was perfused with Krebs Heinseleit buffer at 37deg;C containing 4.7percnt; bovine serum albumin at a constant flow rate of 4 ml per min for 30-60 min with 100percnt; survival of the animals. Tumour vascularization and blood flow were demonstrated using vascular casts and lsqb;51Crrsqb;-microspheres. Following the addition of melphalan (15 or 100micro;gsol;ml), drug concentrations in the perfusate, tissues and systemic circulation were determined using high pressure liquid chromatography (HPLC). Systemic leakage, assessed using lsqb;125lrsqb;albumin and melphalan and detected by a gamma;-counter and HPLC respectively, was0.5percnt;. The melphaian concentration and tissue flow rate in the tumour deposits were 40 and 30percnt; respectively, when compared with the surrounding subcutaneous tissue. At a dose of 15micro;gsol;ml, melphalan caused a reduction in tumour growth after 60 min perfusion, and a significant reduction in tumour size was seen when the melphalan dose was 100micro;gsol;ml. The surviving nude rat model of isolated limb perfusion for recurrent melanoma will allow examination of optimal perfusion conditions, along with the pharmacokinetics, pharmacodynamics and efficacy of melphalan and other drugs.