Immunological, biochemical, ultrastructural, and electrophysiological characteristics of a human glioblastoma-derived cell culture line

1982 ◽  
Vol 56 (1) ◽  
pp. 62-72 ◽  
Author(s):  
Peter McL. Black ◽  
Paul L. Kornblith ◽  
Peter F. Davison ◽  
Theodore M. Liszczak ◽  
Linda P. Merk ◽  
...  
Keyword(s):  
Cell Line ◽  
Surface Membrane ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Human Cell Line ◽  
Resting Membrane Potential ◽  
Content Type ◽  
Actin Cables ◽  
Fine Print

✓ This report presents the results of a study using multiple techniques of the established human cell line, LM, which has been developed in culture medium from a patient with a right temporoparietal glioblastoma. This cell line has human subtetraploid karyotype and has several features of a transformed line in culture. These include continuous propagation for 10 years, ability to form tumor nodules when transplanted into immunologically suppressed hamsters, and pleomorphic appearance. Ultrastructurally, it is characterized by multiple nuclei, few actin cables, and numerous surface-membrane microvilli, as well as abundant 9- to 10-nm cystoplasmic filaments. By its immunological reactivity, the line can be shown to contain glial fibrillary acidic protein at low levels, consistent with its glial origin and continued nature. Dibutyryl cyclic adenosine monophosphate (db-cAMP) induces formation of long astrocytic-like processes as well. Its membrane electrical characteristics include a low resting membrane potential and short time constant. Used in a microtiter antiglioma antibody cytotoxicity assay, LM yields a positive reaction to antibodies in the sera of 80% of patients with astrocytomas and only 9% of normal blood-bank donors, suggesting that it shares common antigens with other astrocytic tumor lines. The varied characteristics of this glioblastoma-derived line emphasize the “multiforme” nature of the neoplasm and suggest that for characterization of any such line, multiple parameters are necessary to allow comparison with other long-term glioblastoma lines in the literature. The usefulness of the LM line in in vitro cell biological, immunological, chemotherapeutic, and radiobiological studies of gliomas makes such efforts very worthwhile.

Immune Competence of Newborn Lymphocytes

PEDIATRICS ◽  
1980 ◽  
Vol 65 (3) ◽  
pp. 491-496
Author(s):  
Zeev T. Handzel ◽  
Stanley Levin ◽  
Zippora Dolphin ◽  
Menachem Schlesinger ◽  
Thalia Hahn ◽  
...  
Keyword(s):  
Cell Function ◽  
Surface Membrane ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Rosette Formation ◽  
Thymidine Uptake ◽  
Resting Cells ◽  
Normal Children ◽  
Immune Competency

The immune competency of peripheral blood (cord) lymphocytes of newborn infants has been investigated by a battery of assays in vitro. Number of T cells was enumerated by E-rosette formation and a cytotoxicity assay by using an anti-human-thymocyte antiserum. Lymphokine production as an indicator of T cell function was evaluated by leukocyte migration inhibition factor production following stimulation with various mitogens, and "viral" and "immune" interferon production in response to stimulation with polyriboinosinic -cytidilic acid and phytohemagglutinin, respectively. Ability to respond to mitogens was also tested by means of 3H-thymidine uptake into DNA, and by measuring the early synthesis of protein by lymphocytes by means of 3H-leucine uptake. Lymphocyte cyclic adenosine monophosphate levels in resting cells and after trypsin treatment was also used as a test of cellular competency. Total B cells was evaluated by EAC-rosette formation, and class specific surface membrane immunoglobulin-bearing cells were assayed by the peroxidase immunoenzymatic method. Normal children and adults served as controls. The results indicated that no significant differences in immune competency could be shown between newborns and older people except for lymphocyte cyclic adenosine monophosphate levels which were lower in the newborns, both in resting and in stimulated cells. The relationship between this latter finding and the immune status is as yet not clear. It is concluded that neonatal lymphocytes are essentially immunocompetent, with the expression of their immune capabilities in vivo becoming apparent only after encounter with environmental antigenic stimuli.

Radiosensitization of C6 glioma by thymidine and 41.8°C hyperthermia

1990 ◽  
Vol 72 (5) ◽  
pp. 782-785 ◽  
Author(s):  
Justin D. Cohen ◽  
H. Ian Robins ◽  
Manucher J. Javid
Keyword(s):  
Cell Survival ◽  
Cell Line ◽  
Untreated Control ◽  
Glioma Cell ◽  
Glioma Cell Line ◽  
C6 Glioma ◽  
Content Type ◽  
C6 Glioma Cell ◽  
Fine Print

✓ The cytotoxic, antiproliferative, and radiosensitizing effects of thymidine (a nucleoside metabolite) were studied using the C6 glioma cell line in vitro. Radiosensitization by a combination of thymidine and 41.8°C hyperthermia was also evaluated. Thymidine concentrations above 100 µg/ml completely inhibited C6 proliferation while concentrations of 100 to 1000 µg/ml (for up to 24 hours) decreased C6 cell survival to as little as 7.4% compared to untreated control cells. Radiosensitivity was enhanced by the administration of thymidine alone (400 µg/ml × 24 hours before irradiation); sensitization by 41.8°C hyperthermia alone (1 hour ending immediately before irradiation) was less pronounced. Thymidine and hyperthermia together produced greater radiosensitization than did heat alone or thymidine alone. These data support the further investigation of thymidine as a neuro-oncology radiosensitizer.

The reversal of experimental vasospasm by dibutyryl-3′, 5′-adenosine monophosphate

1973 ◽  
Vol 39 (6) ◽  
pp. 730-734 ◽  
Author(s):  
Eric W. Peterson ◽  
Roger Searle ◽  
Francis F. Mandy ◽  
Richard Leblanc
Keyword(s):  
Subarachnoid Hemorrhage ◽  
Cyclic Amp ◽  
Cerebral Vasospasm ◽  
Basilar Artery ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Postoperative Management ◽  
Dibutyryl Cyclic Amp ◽  
Content Type ◽  
Fine Print

✓ Topical dibutyryl cyclic adenosine monophosphate (AMP) was used to reverse experimental cerebral vasospasm of the basilar artery in the cat. The combination of dibutyryl cyclic AMP and theophylline caused prolonged dilatation of the basilar artery. Dibutyryl cyclic AMP seems to be specific as a topical vasodilator, which may be useful in the postoperative management of subarachnoid hemorrhage.

scholarly journals Glucagon Prevents Cytotoxicity Induced by Methylglyoxal in a Rat Neuronal Cell Line Model

Biomolecules ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 287
Author(s):  
Mohammad Sarif Mohiuddin ◽  
Tatsuhito Himeno ◽  
Yuichiro Yamada ◽  
Yoshiaki Morishita ◽  
Masaki Kondo ◽  
...  
Keyword(s):  
Nervous System ◽  
Cell Line ◽  
Neuronal Cell ◽  
Cyclic Adenosine Monophosphate ◽  
Diabetic Polyneuropathy ◽  
Adenosine Monophosphate ◽  
Protective Effects ◽  
Neuroprotective Effects ◽  
Neuronal Cell Line

Although diabetic polyneuropathy (DPN) is a frequent diabetic complication, no effective therapeutic approach has been established. Glucagon is a crucial hormone for glucose homeostasis but has pleiotropic effects, including neuroprotective effects in the central nervous system. However, the importance of glucagon in the peripheral nervous system (PNS) has not been clarified. Here, we hypothesized that glucagon might have a neuroprotective function in the PNS. The immortalized rat dorsal root ganglion (DRG) neuronal cell line 50B11 was treated with methylglyoxal (MG) to mimic an in vitro DPN model. The cells were cultured with or without glucagon or MG. Neurotoxicity, survival, apoptosis, neurite projection, cyclic adenosine monophosphate (cAMP), and protein kinase A (PKA) were examined. Glucagon had no cytotoxicity and rescued the cells from neurotoxicity. Cell survival was increased by glucagon. The ratio of apoptotic cells, which was increased by MG, was reduced by glucagon. Neurite outgrowth was accelerated in glucagon-treated cells. Cyclic AMP and PKA accumulated in the cells after glucagon stimulation. In conclusion, glucagon protected the DRG neuronal cells from MG-induced cellular stress. The cAMP/PKA pathway may have significant roles in those protective effects of glucagon. Glucagon may be a potential target for the treatment of DPN.

Hypothalamic hypothyroidism and hypogonadism in prolonged traumatic coma

1978 ◽  
Vol 49 (5) ◽  
pp. 650-657 ◽  
Author(s):  
Alan S. Fleischer ◽  
Daniel R. Rudman ◽  
Nettleton S. Payne ◽  
George T. Tindall
Keyword(s):  
Plasma Concentrations ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Thyroid Stimulating Hormone ◽  
Content Type ◽  
Releasing Hormone ◽  
Traumatic Coma ◽  
Hypothalamic Function ◽  
Fine Print ◽  
Stimulating Hormone

✓ Prolonged coma after head trauma is associated with depletion of 3′,5′cyclic adenosine monophosphate (cAMP) in the cerebrospinal fluid (CSF). Because cAMP has previously been implicated in neuroendocrine secretion, this study examines the pituitary-hypothalamic function in 15 adult male patients (to exclude the effects of puberty and menses) with traumatic coma lasting longer than 2 weeks. Ventricular CSF cAMP was measured at 2- to 4-day intervals for 10 to 25 days. Simultaneously, plasma hormone concentrations were also determined. In all 15 cases, CSF cAMP and plasma levels of thyroid-stimulating hormone (TSH), thyroxine (T4), free T4, triiodothyronine (T3), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone became subnormal. In 11 patients whose level of consciousness fluctuated, the reduction in plasma T4 and testosterone were proportional to both severity of coma (r > 0.81, p < 0.05) and depletion of CSF cAMP (r > 0.81, p < 0.05). In four patients who remained deeply comatose for 17 to 25 days, the hypothyroidism and hypogonadism persisted. In six patients who regained consciousness, both endocrine defects improved partially or completely. Injection of 1) thyrotrophic-releasing hormone and 2) gonadotrophic-releasing hormone elicited normal or supernormal increases in plasma concentrations of 1) TSH, and 2) LH and FSH, reduced, respectively, suggesting a suprahypophyseal deficiency. These observations demonstrate that suprahypophyseal hypothyroidism and hypogonadism may occur regularly in patients with traumatic coma lasting longer than 2 weeks.

Sensitization of rat glioblastoma multiforme to cisplatin in vivo following restoration of wild-type p53 function

1998 ◽  
Vol 88 (3) ◽  
pp. 535-540 ◽  
Author(s):  
Oliver Dorigo ◽  
Sally T. Turla ◽  
Svetlana Lebedeva ◽  
Ruth A. Gjerset
Keyword(s):  
Glioblastoma Multiforme ◽  
Cell Line ◽  
Wild Type ◽  
Content Type ◽  
Fischer 344 ◽  
Cisplatin Sensitivity ◽  
Wild Type P53 ◽  
Fine Print

Object. To study the combined potential of wild-type p53 gene transfer and administration of cisplatin for the treatment of glioblastoma multiforme, the authors used the 9L rat glioblastoma cell line, which expresses a mutant p53. Methods. Stable expression of wild-type p53 in 9L cells was achieved by transfection of the cells with a wild-type p53—expressing plasmid (pCEP4p53). The resultant cell line, 9LpCEP4p53, was found to be more sensitive to cisplatin treatment in vitro than control (9LpCEP4) cells. The in vitro growth rates of control cells and wild-type p53—modified cells were similar in the absence of cisplatin. Fischer 344 rats were implanted intracerebrally with 9LpCEP4p53 cells and intraperitoneally administered 4 mg/kg cisplatin weekly for 7 weeks. These animals survived significantly longer than animals that were implanted with 9LpCEP4p53 cells but were given no cisplatin treatment. In contrast, concurrent cisplatin treatment provided no benefit for animals implanted with 9LpCEP4 cells. Tumors that developed in animals that had been implanted with 9LpCEP4p53 cells and treated with cisplatin had lost expression of wild-type p53, indicating a correlation between expression of wild-type p53 and cisplatin sensitivity in vivo. Conclusions. The findings of this study suggest that p53-based gene therapy in combination with cisplatin-based chemotherapy may be superior to single-modality treatment in dealing with glioblastoma multiforme.

scholarly journals Secretoneurin stimulates the production and release of luteinizing hormone in mouse LβT2 gonadotropin cells

2011 ◽  
Vol 301 (2) ◽  
pp. E288-E297 ◽  
Author(s):  
E. Zhao ◽  
Judy R. McNeilly ◽  
Alan S. McNeilly ◽  
Reiner Fischer-Colbrie ◽  
Ajoy Basak ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Luteinizing Hormone ◽  
Cell Line ◽  
Mammalian Species ◽  
Mrna Level ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Extracellular Signal ◽  
Underlying Mechanisms

Secretoneurin (SN) is a functional secretogranin II (SgII)-derived peptide that stimulates luteinizing hormone (LH) production and its release in the goldfish. However, the effects of SN on the pituitary of mammalian species and the underlying mechanisms remain poorly understood. To study SN in mammals, we adopted the mouse LβT2 gonadotropin cell line that has characteristics consistent with normal pituitary gonadotrophs. Using radioimmunoassay and real-time RT-PCR, we demonstrated that static treatment with SN induced a significant increment of LH release and production in LβT2 cells in vitro. We found that GnRH increased cellular SgII mRNA level and total SN-immunoreactive protein release into the culture medium. We also report that SN activated the extracellular signal-regulated kinases (ERK) in either 10-min acute stimulation or 3-h chronic treatment. The SN-induced ERK activation was significantly blocked by pharmacological inhibition of MAPK kinase (MEK) with PD-98059 and protein kinase C (PKC) with bisindolylmaleimide. SN also increased the total cyclic adenosine monophosphate (cAMP) levels similarly to GnRH. However, SN did not activate the GnRH receptor. These data indicate that SN activates the protein kinase A (PKA) and cAMP-induced ERK signaling pathways in the LH-secreting mouse LβT2 pituitary cell line.

scholarly journals Adenosine A2A receptor signaling promotes FoxO associated autophagy in chondrocytes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Benjamin Friedman ◽  
Carmen Corciulo ◽  
Cristina M. Castro ◽  
Bruce N. Cronstein
Keyword(s):  
Cell Line ◽  
Metabolic Stress ◽  
Human Cell Line ◽  
Adenosine A2a Receptor ◽  
Autophagic Flux ◽  
A2a Receptor ◽  
Adenosine A2a ◽  
A2ar Agonist

AbstractAutophagy, a homeostatic pathway upregulated during cellular stress, is decreased in osteoarthritic chondrocytes and this reduction in autophagy is thought to contribute to the development and progression of osteoarthritis (OA). The adenosine A2A receptor (A2AR) is a potent anti-inflammatory receptor and deficiency of this receptor leads to the development of OA in mice. Moreover, treatment using liposomally conjugated adenosine or a specific A2AR agonist improved joint scores significantly in both rats with post-traumatic OA (PTOA) and mice subjected to a high fat diet obesity induced OA. Importantly, A2AR ligation is beneficial for mitochondrial health and metabolism in vitro in primary and the TC28a2 human cell line. An additional set of metabolic, stress-responsive, and homeostatic mediators include the Forkhead box O transcription factors (FoxOs). Data has shown that mouse FoxO knockouts develop early OA with reduced cartilage autophagy, indicating that FoxO-induced homeostasis is important for articular cartilage. Given the apparent similarities between A2AR and FoxO signaling, we tested the hypothesis that A2AR stimulation improves cartilage function through activation of the FoxO proteins leading to increased autophagy in chondrocytes. We analyzed the signaling pathway in the human TC28a2 cell line and corroborated these findings in vivo in a metabolically relevant obesity-induced OA mouse model. We found that A2AR stimulation increases activation and nuclear localization of FoxO1 and FoxO3, promotes an increase in autophagic flux, improves metabolic function in chondrocytes, and reduces markers of apoptosis in vitro and reduced apoptosis by TUNEL assay in vivo. A2AR ligation additionally enhances in vivo activation of FoxO1 and FoxO3 with evidence of enhanced autophagic flux upon injection of the liposome-associated A2AR agonist in a mouse obesity-induced OA model. These findings offer further evidence that A2AR may be an excellent target for promoting chondrocyte and cartilage homeostasis.

Cerebrovascular characterization of clazosentan, the first nonpeptide endothelin receptor antagonist clinically effective for the treatment of cerebral vasospasm. Part I: Inhibitory effect on endothelinA receptor—mediated contraction

2005 ◽  
Vol 102 (6) ◽  
pp. 1101-1107 ◽  
Author(s):  
Hartmut Vatter ◽  
Michael Zimmermann ◽  
Veronika Tesanovic ◽  
Andreas Raabe ◽  
Lothar Schilling ◽  
...  
Keyword(s):  
Receptor Antagonist ◽  
Cerebral Vasospasm ◽  
Isometric Force ◽  
Inhibitory Effect ◽  
Affinity Constant ◽  
Dependent Manner ◽  
Content Type ◽  
The Right ◽  
Fine Print

Object. The central role of endothelin (ET)—1 in the development of cerebral vasospasm after subarachnoid hemorrhage is indicated by the successful treatment of this vasospasm in several animal models by using selective ETA receptor antagonists. Clazosentan is a selective ETA receptor antagonist that provides for the first time clinical proof that ET-1 is involved in the pathogenesis of cerebral vasospasm. The aim of the present investigation was, therefore, to define the pharmacological properties of clazosentan that affect ETA receptor—mediated contraction in the cerebrovasculature. Methods. Isometric force measurements were performed in rat basilar artery (BA) ring segments with (E+) and without (E−) endothelial function. Concentration effect curves (CECs) were constructed by cumulative application of ET-1 or big ET-1 in the absence or presence of clazosentan (10−9, 10−8, and 10−7 M). The inhibitory potency of clazosentan was determined by the value of the affinity constant (pA2). The CECs for contraction induced by ET-1 and big ET-1 were shifted to the right in the presence of clazosentan in a parallel dose-dependent manner, which indicates competitive antagonism. The pA2 values for ET-1 were 7.8 (E+) and 8.6 (E−) and the corresponding values for big ET-1 were 8.6 (E+) and 8.3 (E−). Conclusions. The present data characterize clazosentan as a potent competitive antagonist of ETA receptor—mediated constriction of the cerebrovasculature by ET-1 and its precursor big ET-1. These functional data may also be used to define an in vitro profile of an ET receptor antagonist with a high probability of clinical efficacy.

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