Stem cell studies of human malignant brain tumors

1985 ◽  
Vol 63 (3) ◽  
pp. 426-432 ◽  
Author(s):  
Bertrand Pertuiset ◽  
Dolores Dougherty ◽  
Carlos Cromeyer ◽  
Takao Hoshino ◽  
Mitchel Berger ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Brain Tumors ◽  
Tritiated Thymidine ◽  
Early Passage ◽  
Cell Cycle Time ◽  
Content Type ◽  
Cell Kill ◽  
Fine Print

✓ The proliferation kinetics were studied in early-passage cultures of cells from 13 human malignant brain tumors and two specimens of normal brain under conditions similar to those used in clonogenic cell-survival studies. Autoradiography was performed in all but four cases to estimate the fraction of cells actively replicating deoxyribonucleic acid (DNA), the approximate cell cycle time, and the effect of low-dose tritiated thymidine on cell proliferation. The mean tumor cell doubling time (TD) was 53 hours for five glioblastomas, 46 hours for two ependymomas, and 83 hours for two medulloblastomas. A gliosarcoma grew fastest (TD = 22 hours) in culture and a pilocytic astrocytoma grew slowest (TD = 144 hours). The approximate cell cycle time ranged from 1 to 2.5 days for all tumors tested. This suggests that chemotherapeutic agents that predominantly kill proliferating cells should be administered in vitro for at least 2 to 2.5 days to achieve maximum cell kill. The approximate growth fraction ranged from 0.65 to 0.96 for all tumors except for the two medulloblastomas and the pilocytic astrocytoma, which had growth fractions of 0.34 and 0.35, respectively. Most laboratories investigating the chemosensitivity of primary or early-passage human tumor cells require that 40% to 70% of cells be killed to consider a drug active in vitro. The results of this study suggest that the cell-cycle-specific agents cannot achieve a high enough cell kill to be considered active for some tumors that grow slowly in culture. An estimate of the in vitro growth rate is necessary to reliably interpret cell-survival results with such agents. Tritiated thymidine appeared to slow cell proliferation in some of the cultures, presumably as a result of radiation-induced DNA damage caused by tritium that had been incorporated into DNA. The degree to which cell growth was slowed in individual tumors correlated with the patient's clinical response to radiation therapy and postoperative survival time.

scholarly journals Differences in cell cycle characteristics among patients with acute nonlymphocytic leukemia

Blood ◽  
1987 ◽  
Vol 69 (6) ◽  
pp. 1647-1653 ◽  
Author(s):  
A Raza ◽  
Y Maheshwari ◽  
HD Preisler
Keyword(s):  
Cell Cycle ◽  
Tritiated Thymidine ◽  
S Phase ◽  
Total Cell ◽  
Acute Nonlymphocytic Leukemia ◽  
Cell Cycle Time ◽  
Myeloid Leukemias ◽  
History Of

The proliferative characteristics of myeloid leukemias were defined in vivo after intravenous infusions of bromodeoxyuridine (BrdU) in 40 patients. The percentage of S-phase cells obtained from the biopsies (mean, 20%) were significantly higher (P = .00003) than those determined from the bone marrow (BM) aspirates (mean, 9%). The post- BrdU infusion BM aspirates from 40 patients were incubated with tritiated thymidine in vitro. These double-labeled slides were utilized to determine the duration of S-phase (Ts) in myeloblasts and their total cell cycle time (Tc). The Ts varied from four to 49 hours (mean, 19 hours; median, 17 hours). Similarly, there were wide variations in Tc of individual patients ranging from 16 to 292 hours (mean, 93 hours; median, 76 hours). There was no relationship between Tc and the percentage of S-phase cells, but there was a good correlation between Tc and Ts (r = .8). Patients with relapsed acute nonlymphocytic leukemia (ANLL) appeared to have a longer Ts and Tc than those studied at initial diagnosis. A subgroup of patients at either extreme of Tc were identified who demonstrated clinically documented resistance in response to multiple courses of chemotherapy. We conclude that Ts and Tc provide additional biologic information that may be valuable in understanding the variations observed in the natural history of ANLL.

Immunobiology of primary intracranial tumors

1981 ◽  
Vol 54 (3) ◽  
pp. 331-337 ◽  
Author(s):  
William H. Brooks ◽  
Robert B. Latta ◽  
M. Stephen Mahaley ◽  
Thomas L. Roszman ◽  
Lynn Dudka ◽  
...  
Keyword(s):  
Brain Tumors ◽  
Linear Combination ◽  
Tumor Recurrence ◽  
Intracranial Tumors ◽  
Content Type ◽  
Sequential Analyses ◽  
Fine Print ◽  
Host Immunocompetence

✓ Long-term assessment of general host immunocompetence of patients with primary malignant brain tumors indicates that although isolated determinations of nonspecific responsiveness are not clinically useful, sequential analyses utilizing a linear combination of in vitro lymphocyte probes are capable of predicting tumor recurrence prior to clinical deterioration.

CELL PROLIFERATION IN THE PREPUBERTAL MALE RAT ADRENAL CORTEX: AN AUTORADIOGRAPHIC STUDY

1971 ◽  
Vol 49 (4) ◽  
pp. 599-609 ◽  
Author(s):  
N. A. WRIGHT
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Adrenal Cortex ◽  
Tritiated Thymidine ◽  
Autoradiographic Study ◽  
Male Rats ◽  
Male Rat ◽  
General Increase ◽  
Cortical Cells ◽  
Cell Cycle Time

SUMMARY On the basis of labelling indices measured with tritiated thymidine at intervals throughout its thickness, the adrenal cortex of prepubertal male rats has been divided into four compartments. These are called the glomerular, proliferative, fascicular and reticular compartments, respectively. Labelling indices measured for each compartment showed highest values in the glomerular and proliferative compartments, with values of 6·73% and 7·09% respectively. The fascicular compartment showed a lower index of 3·16% while the reticular compartment gave the lowest value of 1·15%. These differences are further reflected in measurements of the mitotic indices for each compartment. The phases of the cell cycle have been measured by pulse-chase analysis in each compartment, and all phases estimated showed an increase in duration as the inner compartments were approached. The duration of interphase DNA synthesis (ts) was found to be shortest in the glomerular and proliferative compartments, with values of 7·45 and 7·73 h, respectively. The fascicular compartment showed lengthening of ts to 8·56 h, and the reticular compartment gave the highest value of 9·21 h. Similarly, the values obtained for G2 (the post-DNA synthetic interval) and tm (the duration of mitosis), and a calculated value of the cell cycle time all showed a general increase in duration from the outer to the inner compartments. The relation of these results to theories of adrenocortical cytogenesis is discussed, and it is suggested that the differences in cell cycle components can best be explained by the inward migration of cortical cells from the outer compartments.

Treatment of murine primary brain tumors with systemic interleukin-2 and tumor-infiltrating lymphocytes

1992 ◽  
Vol 76 (3) ◽  
pp. 513-519 ◽  
Author(s):  
Stephen C. Saris ◽  
Paul Spiess ◽  
Daniel M. Lieberman ◽  
Shan Lin ◽  
Stuart Walbridge ◽  
...  
Keyword(s):  
Brain Tumors ◽  
Interleukin 2 ◽  
Tumor Infiltrating Lymphocytes ◽  
Similar Response ◽  
Content Type ◽  
Primary Brain Tumors ◽  
Infiltrating Lymphocytes ◽  
The Brain ◽  
Fine Print

✓ Methods have recently been described for the isolation and expansion of lymphocytes that have trafficked into animal and human tumors. These CD8-positive tumor-infiltrating lymphocytes (TIL's) have exceptional trafficking ability to, and efficacy against, tumor targets in extracranial sites. Prior to Phase I clinical trials for patients with gliomas, adoptive immunotherapy with TIL's was studied in a mouse model of primary brain tumors to determine if intracerebral tumors have a similar response. Glioma 261 (GL261) tumors were grown in the subcutaneous space of C57BL/6 mice. After enzymatic digestion, the cells were incubated in vitro with interleukin-2 (IL-2) until a confluent population of T lymphocytes was present. The in vitro efficacy of these TIL's was tested against fresh GL261 targets with a chromium release assay; the in vivo efficacy was tested against GL261 tumors in the liver and against irradiated and nonirradiated GL261 tumors in the brain. Mice received one of the following: intraperitoneal saline; intraperitoneal IL-2 (7500 to 50,000 U three times daily for 5 days); IL-2 plus intravenous TIL's (1 to 3 × 107 cells); 10 Gy cranial irradiation; irradiation plus IL-2; or irradiation plus IL-2 plus TIL's. The TIL preparation killed 77% of tumor targets in 4 hours at an effector:target ratio of 100:1. In animals with GL261 tumors in the liver, at 2 weeks there were 93 ± 37, 128 ± 45, and 21 ± 14 liver metastases in the control, IL-2, and IL-2 plus TIL groups, respectively. However, in animals with GL261 tumors in the brain, no treatment group had an increased survival rate compared to the control group. It is concluded that, although TIL and IL-2 immunotherapy can be used effectively to treat brain tumors in vitro and at sites outside the central nervous system, it is ineffective against the same type of tumor in the brain. Different methods of delivery or different combinations of these immunomodulators may be more effective; however, based on these findings, treatment of patients with IL-2 and TIL cannot be recommended until efficacy has been demonstrated in an animal model.

scholarly journals Simultaneous immunohistochemical detection of IUdR and BrdU infused intravenously to cancer patients.

1991 ◽  
Vol 39 (4) ◽  
pp. 407-412 ◽  
Author(s):  
M A Miller ◽  
C M Mazewski ◽  
N Yousuf ◽  
Y Sheikh ◽  
L M White ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Solid Tumors ◽  
Bone Marrow Cells ◽  
Tritiated Thymidine ◽  
Culture Cell ◽  
S Phase ◽  
Tissue Culture Cell ◽  
Cell Cycle Time

Cell cycle kinetics of solid tumors in the past have been restricted to an in vitro labeling index (LI) measurement. Two thymidine analogues, bromodeoxyuridine (BrdU) and iododeoxyuridine (IUdR), can be used to label S-phase cells in vivo because they can be detected in situ by use of monoclonal antibodies (MAb) against BrdU (Br-3) or IUdR (3D9). Patients with a variety of solid tumors (lymphoma, brain, colon cancers) received sequential intravenous IUdR and BrdU. Tumor tissue removed at the end of infusion was embedded in plastic and treated with MAb Br-3 and 3D9 sequentially, using a modification of a previously described method. Clearly single and double labeled cells were visible, which enabled us to determine the duration of S-phase (Ts) and the total cell cycle time (Tc), in addition to the LI in these tumors. Detailed control experiments using tissue culture cell lines as well as bone marrow cells from leukemic patients are described, including the comparison of this double label technique with our previously described BrdU-tritiated thymidine technique. We conclude that the two methods are comparable and that the IUdR/BrdU method permits rapid and reliable cell cycle measurements in solid tumors.

Dynamic determination of human glioma invasion in vitro

1998 ◽  
Vol 89 (3) ◽  
pp. 441-447 ◽  
Author(s):  
Svein J. T. Nygaard ◽  
Hans K. R. Haugland ◽  
Ole Didrik Laerum ◽  
Morten Lund-Johansen ◽  
Rolf Bjerkvig ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Tumor Cell ◽  
Brain Tissue ◽  
Normal Brain ◽  
Tumor Spheroids ◽  
Normal Brain Tissue ◽  
Content Type ◽  
Biopsy Specimens ◽  
Fine Print

Object. The goal of this study was to evaluate whether there is any relationship between survival of patients with brain tumor and tumor proliferation or tumor invasion in vitro. Methods. Samples of freshly resected brain tumors from 14 patients with glioblastoma multiforme (GBM) were directly grown as three-dimensional multicellular spheroids. The tumor spheroids were cocultured with fetal rat brain cell aggregates (BCAs), used to represent an organotypical normal brain tissue model. Before the coculture, the tumor spheroids and the BCAs were stained with two different carbocyanine dyes, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) and 3,3′-dioctadecycloxacarbocyanine perchlorate (DiO), respectively. During the coculture, confocal laser scanning microscopy allowed a sequential analysis of tumor cell invasion by visualizing dynamic aspects of the invasive process. Single cocultures were examined at three different time points (24, 48, and 96 hours). During the observation period there was a change in the structural morphology of the cocultures, with a progressive decrease in BCA volume. Furthermore, the scanning confocal micrographs revealed a bidirectional movement of tumor cells and normal cells into brain and tumor tissue, respectively. It is also shown that there is a considerable variation in the rate of BCA destruction in cocultures of glioma spheroids generated directly from biopsy specimens. This variation is seen both between spheroids generated from the same biopsy as well as between spheroids that are grown from different biopsy specimens. Cell proliferation measured by Ki-67 immunohistochemical analysis of biopsy samples obtained in the same patients revealed a correlation between tumor cell proliferation and tissue destruction of the BCAs, as determined by a reduction in BCA volume (p = 0.0338). No correlation was found when survival was related to the same parameters (p > 0.05). Conclusions. The present work provides a model for quick and efficient assessment of dynamic interactions between tumor and normal brain tissue shortly after surgery.

Correlation of in vitro bromodeoxyuridine labeling index and DNA aneuploidy with survival or recurrence in brain-tumor patients

1990 ◽  
Vol 73 (3) ◽  
pp. 396-400 ◽  
Author(s):  
Takafumi Nishizaki ◽  
Tetsuji Orita ◽  
Koji Kajiwara ◽  
Norio Ikeda ◽  
Noboru Ohshita ◽  
...  
Keyword(s):  
Brain Tumor ◽  
Brain Tumors ◽  
Survival Data ◽  
Patient Survival ◽  
Labeling Index ◽  
Content Type ◽  
Dna Aneuploidy ◽  
Labeling Method ◽  
Fine Print

✓ There are no previous reports correlating the in vitro bromodeoxyuridine (BUdR) labeling index (LI) with the clinical outcome in patients with brain tumors. The reliability of the LI as a predictor of patient survival or recurrence was examined in this study of 66 human brain tumors (19 gliomas, 18 meningiomas, and 29 others). Anti-BUdR staining was performed on surgically extirpated tumor tissue that had been treated with BUdR as previously described. Correlation of the BUdR LI with patient survival or tumor recurrence rate was carried out by the method of Kaplan and Meier. Deoxyribonucleic acid (DNA) aneuploidy was estimated in 52 cases. The results of this study indicate that BUdR LI values correlated well with the clinical course of patients with brain tumor. In comparison with patients with higher LI's, there was both a significantly higher survival rate for tumors other than meningiomas and a higher recurrence-free rate for meningiomas in patients with LI's of less than 4% and 1%, respectively. Although there was a tendency for patients without tumor aneuploidy to show better survival data than the others, no statistical difference was observed. These results suggest that the in vitro BUdR labeling method is reliable for prediction of a patient's prognosis, whereas prognosis on the basis of DNA aneuploidy alone is uncertain.

scholarly journals Locomotion and proliferation of glioblastoma cells in vitro: statistical evaluation of videomicroscopic observations

2000 ◽  
Vol 92 (3) ◽  
pp. 428-434 ◽  
Author(s):  
Balás Hegedüs ◽  
András Czirók ◽  
Ilona Fazekas ◽  
Tamás Bábel ◽  
Emília Madarász ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
Cycle Time ◽  
Average Velocity ◽  
Statistical Evaluation ◽  
Glioblastoma Cells ◽  
Cell Cycle Time ◽  
Content Type ◽  
Cell Locomotion ◽  
Fine Print

Object. The motility and doubling of human glioblastoma cells were investigated by means of statistical evaluation of large sets of data obtained using computer-aided videomicroscopy.Methods. Data were obtained on cells in four established glioblastoma cell lines and also on primary tumor cells cultured from fresh surgical samples. Growth rates and cell cycle times were measured in individual microscopic fields. The averages of cell cycle time and the duplication time for the recorded cell populations were 26.2 ± 5.6 hours and 38 ± 4 hours, respectively. With these parameters, no significant differences among the cell lines were revealed. Also, there was no correlation in the cell cycle time of a parent cell and its progeny in any of the cultures.Statistical analysis of cell locomotion revealed an exponential distribution of cell velocities and strong fluctuations in individual cell velocities across time. The average velocity values ranged from 4.2 to 27.9 µm/hour. In spite of the uniform histopathological classification of the four tumors, each cell line produced by these tumors displayed distinct velocity distribution profiles and characteristic average velocity values. A comparison of recently established primary cultures with cell lines that had propagated multiple times indicated that cells derived from different tumors sustain their characteristic locomotor activity after several passages.Conclusions. It can be inferred from the data that statistical evaluation of physical parameters of cell locomotion can provide additional tools for tumor diagnosis.

Immunohistochemical demonstration of DNA polymerase α in human brain-tumor cells

1990 ◽  
Vol 72 (2) ◽  
pp. 268-272 ◽  
Author(s):  
Katsuzo Kunishio ◽  
Nobuya Mishima ◽  
Takashi Matsuhisa ◽  
Kazuyuki Tsuno ◽  
Nobuhiko Matsumi ◽  
...  
Keyword(s):  
Brain Tumor ◽  
Brain Tumors ◽  
Human Brain ◽  
Dna Polymerase ◽  
Ki 67 ◽  
Content Type ◽  
Brain Tumor Cells ◽  
Fine Print

✓ The proliferative capacity of brain-tumor cells was analyzed in vitro and in situ using monoclonal antibody (MAb) against deoxyribonucleic acid (DNA) polymerase α. For the in vitro studies, two cultured human glioma cell lines were investigated using MAb against DNA polymerase α, the MAb Ki-67, a serum against proliferating cell nuclear antigen (PCNA/cyclin), bromodeoxyuridine (BUdR), and an anti-BUdR MAb. During exponential growth of the cells, the percentage of polymerase α-positive cells (the “polymerase α score”) ranged from 72.0% to 77.1%, the Ki-67-positive cells (the “Ki-67 score”) ranged from 43.4% to 59.4%, the PCNA/cyclin-positive cells from 30.9% to 41.4%, and the BUdR labeling index from 28.6% to 39.3%. For the in situ studies, tissue from 60 human brain tumors and from two normal human brains was investigated and the polymerase α scores and Ki-67 scores were compared. In normal brain tissue, no immunostaining was found by either method. In brain tumors, both the polymerase α scores and the Ki-67 scores correlated with the histological grade of malignancy. Polymerase α scores were generally higher than Ki-67 scores in the same specimen, especially in malignant brain tumors. These findings suggest that immunostaining of DNA polymerase α is a convenient and important new method by which to estimate the cellular proliferation rate of brain tumors. Polymerase α scores may be closer to the growth fraction of the individual tumor than the MAb Ki-67 or other scores.

scholarly journals Differences in cell cycle characteristics among patients with acute nonlymphocytic leukemia

Blood ◽  
1987 ◽  
Vol 69 (6) ◽  
pp. 1647-1653 ◽  
Author(s):  
A Raza ◽  
Y Maheshwari ◽  
HD Preisler
Keyword(s):  
Cell Cycle ◽  
Tritiated Thymidine ◽  
S Phase ◽  
Total Cell ◽  
Acute Nonlymphocytic Leukemia ◽  
Cell Cycle Time ◽  
Myeloid Leukemias ◽  
History Of

Abstract The proliferative characteristics of myeloid leukemias were defined in vivo after intravenous infusions of bromodeoxyuridine (BrdU) in 40 patients. The percentage of S-phase cells obtained from the biopsies (mean, 20%) were significantly higher (P = .00003) than those determined from the bone marrow (BM) aspirates (mean, 9%). The post- BrdU infusion BM aspirates from 40 patients were incubated with tritiated thymidine in vitro. These double-labeled slides were utilized to determine the duration of S-phase (Ts) in myeloblasts and their total cell cycle time (Tc). The Ts varied from four to 49 hours (mean, 19 hours; median, 17 hours). Similarly, there were wide variations in Tc of individual patients ranging from 16 to 292 hours (mean, 93 hours; median, 76 hours). There was no relationship between Tc and the percentage of S-phase cells, but there was a good correlation between Tc and Ts (r = .8). Patients with relapsed acute nonlymphocytic leukemia (ANLL) appeared to have a longer Ts and Tc than those studied at initial diagnosis. A subgroup of patients at either extreme of Tc were identified who demonstrated clinically documented resistance in response to multiple courses of chemotherapy. We conclude that Ts and Tc provide additional biologic information that may be valuable in understanding the variations observed in the natural history of ANLL.

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