Animal model of intramedullary spinal cord glioma using human glioblastoma multiforme neurospheres.
ABSTRACT Advances in the diagnosis and management of patients with spinal cord tumors have been limited because of the rarity of the disease and the limitations of current animal models for spinal cord glioma. The ideal spinal cord tumor model would possess a number of characteristics, including the use of human glioma cells that capture the growth pattern and local invasive nature of their human counterpart. In this study, the authors' goal was to develop a novel spinal cord tumor model using a human neurosphere cell line.
Eighteen female athymic rats were randomized into 3 experimental groups. Animals in the first group (6 rats) received a 3-ml intramedullary injection containing DMEM and were used as controls. Animals in the second group (6 rats) received a 3-ml intramedullary injection containing 100,000 glioblastoma multiforme (GBM) neurosphere cells in 3 ml DMEM. Animals in the third group (6 rats) received a 3-ml intramedullary injection containing 9L gliosarcoma cells in 3 ml DMEM. Functional testing of hindlimb strength was assessed using the Basso-Beattie-Bresnahan (BBB) scale. Once the functional BBB score of an animal was less than or equal to 5 (slight movement of 2 joints and extensive movement of the third), euthanasia was performed.
Animals in the GBM neurosphere group had a mean survival of 33.3 ± 2.0 days, which was approximately twice as long as animals in the 9L gliosarcoma group (16.3 ± 2.3 days). There was a significant difference between survival of the GBM neurosphere and 9L gliosarcoma groups (p < 0.001). None of the control animals died (p < 0.001 for GBM neurosphere group vs controls and 9L vs controls). Histopathological examination of the rats injected with 9L gliosarcoma revealed that all animals developed highly cellular, well-circumscribed lesions causing compression of the surrounding tissue, with minimal invasion of the surrounding gray and white matter. Histopathological examination of animals injected with GBM neurospheres revealed that all animals developed infiltrative lesions with a high degree of white and gray matter invasion along with areas of necrosis.
The authors have established a novel animal model of spinal cord glioma using neurospheres derived from human GBM. When injected into the spinal cords of athymic nude rats, neurospheres gave rise to infiltrative, actively proliferating tumors that were histologically identical to spinal cord glioma in humans. On the basis of their results, the authors conclude that this is a reproducible animal model of high-grade spinal cord glioma based on a human GBM neurosphere line. This model represents an improvement over other models using nonhuman glioma cell lines. Novel therapeutic strategies can be readily evaluated using this model.
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ABSTRACT: From January 1962 to May 1979, 23 patients with biopsy-proved intramedullary spinal cord tumors were treated initially with total resection, subtotal resection, irradiation, or subtotal resection and irradiation. Local control was achieved in 1/2 patients after total resection, 1/3 after subtotal resection, 8/9 after subtotal resection and radiation therapy, and 5/8 after radiation therapy alone. The patients with ependymomas exhibited a radiation dose-response relationship; of eight patients followed five or more years postirradiation, local control was achieved in 2/3 with time dose fraction (TDF) less than 55, 2/3 with TDF 55-65, and 2/2 with TDF greater than 65. The actuarial 5- and 10-year survival rates were 58% and 23% for astrocytoma, and 100% and 73% for patients with ependymoma, respectively. Neurological deficits improved or became totally normal after initial irradiation. Patterns of failure, management of recurrences, and radiotherapeutic techniques and dose recommendations are discussed.Radiology 06/1980; 135(2):473-9. DOI:10.1148/radiology.135.2.7367644 · 6.21 Impact Factor
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ABSTRACT: Intramedullary spinal cord tumors are difficult lesions to treat given their recurrence rate and limited treatment options. The absence of an adequate animal model, however, has hindered the development of new treatment paradigms. In this study, we describe the technique for intramedullary injection of two experimental rodent gliomas (9L and F98) and present the methodology for functional and histopathological analysis of tumor progression. F344 rats (n = 24) were randomized into three groups. Group 1 (n = 8) received a 5 microl intramedullary injection of Dulbecco's modified Eagle medium, Group 2 received a 5 microl intramedullary injection of 9L gliosarcoma (100,000) cells, and Group 3 received a 5 microl intramedullary injection of F98 glioma (100,000) cells. The animals were anesthetized, a 2 cm incision was made in the dorsal mid-thoracic region, and the spinous process of the T5 vertebrae was removed to expose the intervertebral space. The ligamentum flavum was removed, and an intramedullary injection was made into the spinal cord. The animals were evaluated daily for signs of paralysis using the Basso, Beattie, and Bresnahan scale and sacrificed after the onset of deficits for histopathological analysis. Animals injected with 9L-gliosarcoma had a median onset of hind limb paresis at 12 +/- 2.9 days. Animals injected with F98 glioma had a median onset of hind limb paresis at 19 +/- 3 days. Animals injected with Dulbecco's modified Eagle medium did not show neurological deficits. Hematoxylin-eosin cross sections confirmed the presence of intramedullary 9L and F98 tumor invading the spinal cord. Control animals had no significant histopathological findings. Animals injected with 9L or F98 consistently developed hind limb paresis in a reliable and reproducible manner. The progression of neurological deficits is similar to that seen in patients with intramedullary spinal cord tumors. These findings suggest that this model mimics the behavior of intramedullary spinal cord tumors in humans and may be used to examine the efficacy of new treatment options for both low- and high-grade intramedullary tumors.Neurosurgery 08/2006; 59(1):193-200; discussion 193-200. DOI:10.1227/01.NEU.0000219276.44563.DA · 3.03 Impact Factor
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ABSTRACT: Primary spinal cord tumors are rare, and treatment recommendations are therefore difficult. We reviewed a 22-year experience of postoperative radiotherapy for spinal cord tumors to elucidate prognostic factors and recommendations. Twenty-two patients with spinal cord tumors were treated from 1969-1991. Ten patients had ependymomas, of which two were high grade. Twelve had astrocytomas, of which 4 were high grade. Karnofsky status, age, extent of resection, tumor histology, grade, and radiation dose were evaluated, as well as degree of clinical improvement after treatment based on change in Karnofsky status. Ependymomas achieved 100% local control with postoperative radiotherapy. Grade and dose were of indeterminate significance because of these excellent results. High-grade astrocytomas all recurred and caused death. Disease recurred in 1 of 8 patients with low-grade astrocytic or mixed astrocytic tumors. The only prognostic variables of significance were histology, grade, and change in Karnofsky status after treatment. Radiation of primary spinal cord tumors is rare. In nearly all cases, local fields may be used. Improvement in Karnofsky status after radiotherapy may predict better survival. Treatment recommendations for these rare tumors are discussed.Radiation Oncology Investigations 01/1998; 6(6):276-80. DOI:10.1002/(SICI)1520-6823(1998)6:6<276::AID-ROI5>3.0.CO;2-6