Determination of Intracranial Tumor Volumes in a Rodent Brain Using MagneticResonance
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Determination of Intracranial Tumor Volumes in a Rodent Brain Using Magnetic Resonance Imaging
Abstract
The measurement of tumor volumes is a practical andobjective method of assessing the efficacy of a therapeutic agent.However, the relative accuracy of different methods of assessingtumor volume has been unclear. Using 1-weighted, gadoliniumenhancedmagnetic resonance Imaging ( 1-MRI), Evans Blue infusionand histology we measured intracranial tumor volumes ina rodent brain tumor model (RT2) at days 10, 16 and 18 after implantationof cells in the caudate putamen. There is a good correlationbetween tumor volumes comparing 1-MRI and Evans Blue( 2 = 0 99), 1-MRI and Histology ( 2 = 0 98) and histologyand Evans Blue ( 2 = 0 93). Each of these methods is reliable inestimating tumor volumes in laboratory animals. There was significantuptake of gadolinium and Evans Blue in the tumor suggestinga wide disruption of the blood-brain barrier.
Index Terms—Blood-brain barrier, Evans Blue, magnetic resonanceimaging (MRI), tumor volumes.
I. INTRODUCTION
THE MEASUREMENT of tumor volumes in the brainprovides an objective method of studying the effect oftherapeutic agents both in laboratory animals and in humans.There are a number of intracranial rodent tumor modelspresently being studied for the assessment of the efficacy ofchemotherapeutic agents [1]. We used the RT2 cell line as theintracranial tumor model. The RT2 cell line is derived froman Avian Sarcoma Virus (Schmidt–Ruppin subgroup) inducedanaplastic rat cerebral glioma [2], [3]. This intracerebral gliomamodel has proven useful in the study of radiotherapeutic,chemotherapeutic, and immunotherapeutic approaches for thecontrol of intracranial glioma growth [4], [5]. The histologicalappearance of RT2 gliomas is very similar but not identical to the appearance of spontaneously occurring human gliomas.This includes large numbers of mitotic figures, nuclear pleomorphism,vascular proliferation, hemorrhage, and necrosis.Although the margins of the RT2 tumors in gross and histologicsections appear relatively discrete, these tumors also demonstratefeatures of infiltration into surrounding brain, which isthe sine-qua-non of human malignant gliomas. Nevertheless,as with human gliomas, growth of the intracerebral mass isa direct result of tumor cell proliferation and is the primarymechanism of morbidity in the animals harboring them. Inour tumor model the rats demonstrate signs and symptomsof raised intracranial pressure due to mass effect from day12 after implantation of tumor cells, and rapidly deterioratearound day 20, at which time they are sacrificed. In order tocompare methods, we measured tumor volumes at 10, 16, and18 days following tumor implantation, utilizing three differenttechniques in each of multiple animals at each time point.-MRI is a dynamic and practical method for the estimationof intracranial tumor volumes in both small and larger animals[6]–[9]. Here, we describe two other techniques usingEvans Blue infusion and histological sections, respectively, toestimate tumor volumes and study the relationship between thethree techniques. These results represent the first collective intercomparisonof all three techniques and they provide a usefulcalibration of each methodology against the other, with potentiallysignificant clinical ramifications for tumor therapies.II. MATERIALS AND METHODSFifteen female Fischer rats were used in this study, with fiverats in each group. The first groupwas sacrificed at 10 days posttumorinnoculation, the second group at 16 days, and the third at18 days. A larger number of animals would have been impracticalto handle logistically, and five per group could still leadto statistically meaningful results. The rats were anesthetizedwith sodium pentobarbital 30–50 mgm/kg (i.p.) body weightand mounted on a Kopf stereotactic apparatus (David Kopf Instruments,Tujunga, CA). The right cerebral hemisphere was inoculatedwith 10 RT2 cells suspended in 5 l of media usinga Hamilton syringe via a burr hole. The landmarks for the injectioninto the caudate putamen were 3.5 and 1 mm to the rightand behind the bregma, respectively, and 4.5mmdeep to the pialsurface. These landmarks were chosen to minimize the growthof the tumor close to the surface of the brain and also to avoidimplantation of tumor cells into the lateral ventricle


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