scholarly journals Erythropoietin production in a primary culture of human renal carcinoma cells maintained in nude mice

Blood ◽  
1984 ◽  
Vol 63 (4) ◽  
pp. 828-835 ◽  
Author(s):  
M Hagiwara ◽  
IL Chen ◽  
R McGonigle ◽  
B Beckman ◽  
FH Kasten ◽  
...  
Keyword(s):  
Nude Mice ◽  
Renal Carcinoma ◽  
Cultured Cells ◽  
Culture Media ◽  
Primary Cultures ◽  
Rabbit Antiserum ◽  
Biologically Active ◽  
Intense Staining ◽  
Human Renal Carcinoma ◽  
Spent Media

Abstract The present studies report erythropoietin (Ep) production in primary cultures of a human renal carcinoma from a patient with erythrocytosis that has been serially transplanted to BALB/c nude mice. The levels of erythropoietin in the culture media were estimated using the exhypoxic polycythemic mouse assay (EHPCMA), fetal mouse liver erythroid colony- forming technique (FMLC), and a radioimmunoassay (RIA). The spent culture media of the exponentially growing cells contained less than 10 mU/ml of Ep measured by RIA. However, after the cells became confluent, Ep levels (RIA) in the spent media showed a marked increase to approximately 300 mU/ml. Ep levels estimated using the FMLC and EHPCMA were approximately 2/3 and 1/10, respectively, of those measured by RIA. Rabbit antiserum to highly purified human urinary Ep (70,400 U/mg protein) was utilized for immunocytochemical (peroxidase-antiperoxidase method) localization of Ep in the cultured cells. Very few of the cells in exponential growth exhibited Ep-like immunoreactivity, whereas intense Ep-like immunoreactivity was observed in the cytoplasm of the cells maintained in culture for a prolonged period after reaching confluency. The most intense staining was observed in some of the cells forming domes. The domes developed after the cells reached confluency, and their numbers increased with increasing time in confluent culture, in parallel with the increase in Ep levels in the spent media. This primary cell culture system of a renal cell carcinoma maintained in nude mice, which produces immunologically and biologically active Ep, may provide a useful model for studies of the mechanism of Ep production.

scholarly journals Erythropoietin production in a primary culture of human renal carcinoma cells maintained in nude mice

Blood ◽  
1984 ◽  
Vol 63 (4) ◽  
pp. 828-835 ◽  
Author(s):  
M Hagiwara ◽  
IL Chen ◽  
R McGonigle ◽  
B Beckman ◽  
FH Kasten ◽  
...  
Keyword(s):  
Nude Mice ◽  
Renal Carcinoma ◽  
Cultured Cells ◽  
Culture Media ◽  
Primary Cultures ◽  
Rabbit Antiserum ◽  
Biologically Active ◽  
Intense Staining ◽  
Human Renal Carcinoma ◽  
Spent Media

The present studies report erythropoietin (Ep) production in primary cultures of a human renal carcinoma from a patient with erythrocytosis that has been serially transplanted to BALB/c nude mice. The levels of erythropoietin in the culture media were estimated using the exhypoxic polycythemic mouse assay (EHPCMA), fetal mouse liver erythroid colony- forming technique (FMLC), and a radioimmunoassay (RIA). The spent culture media of the exponentially growing cells contained less than 10 mU/ml of Ep measured by RIA. However, after the cells became confluent, Ep levels (RIA) in the spent media showed a marked increase to approximately 300 mU/ml. Ep levels estimated using the FMLC and EHPCMA were approximately 2/3 and 1/10, respectively, of those measured by RIA. Rabbit antiserum to highly purified human urinary Ep (70,400 U/mg protein) was utilized for immunocytochemical (peroxidase-antiperoxidase method) localization of Ep in the cultured cells. Very few of the cells in exponential growth exhibited Ep-like immunoreactivity, whereas intense Ep-like immunoreactivity was observed in the cytoplasm of the cells maintained in culture for a prolonged period after reaching confluency. The most intense staining was observed in some of the cells forming domes. The domes developed after the cells reached confluency, and their numbers increased with increasing time in confluent culture, in parallel with the increase in Ep levels in the spent media. This primary cell culture system of a renal cell carcinoma maintained in nude mice, which produces immunologically and biologically active Ep, may provide a useful model for studies of the mechanism of Ep production.

Pathogenesis of Hypercalcemia in Nude Mice Bearing a Human Renal Carcinoma*

Endocrinology ◽  
1986 ◽  
Vol 119 (1) ◽  
pp. 303-310 ◽  
Author(s):  
G. J. STREWLER ◽  
T. J. WRONSKI ◽  
B. P. HALLORAN ◽  
S. C. MILLER ◽  
S. C. LEUNG ◽  
...  
Keyword(s):  
Nude Mice ◽  
Renal Carcinoma ◽  
Human Renal Carcinoma

Cyst Fluids of Malignant Human Brain Tumors Contain Substances That Stimulate the Growth of Cultured Human Gliomas of Various Histological Type

Neurosurgery ◽  
1989 ◽  
Vol 25 (2) ◽  
pp. 196-201 ◽  
Author(s):  
Manfred Westphal ◽  
Hildergard Nausch ◽  
Hans-Dietrich Herrmann
Keyword(s):  
Cell Lines ◽  
Cultured Cells ◽  
Culture Media ◽  
Primary Cultures ◽  
Calf Serum ◽  
Needle Aspiration ◽  
Acoustic Neurinoma ◽  
Response Patterns ◽  
Human Gliomas ◽  
Cyst Fluids

Abstract The contents of 14 cysts that were located within human intracranial tumors were obtained at surgery by needle aspiration. These tumor cyst fluids (TCFs) were mostly derived from glial tumors (10 cases). TCFs from one metastasis from a mammary carcinoma, one cystic meningioma, one hemangioblastoma, and a cystic acoustic neurinoma were also included. These TCFs were added to primary cultures of human gliomas, established human glioma cell lines, and normal human arachnoid cells in culture. The presence of proliferation-promoting factors in all cyst fluids could be demonstrated. On the basis of the response patterns of the cultures, it was possible to distinguish different levels of growth autonomy and growth factor sensitivity among these cultures and to speculate about varying degrees of cellular autocrine activation. The TCFs appear to contain factors that are not normally present in fetal calf serum, which is a regular constituent of most cell culture media. Some primary cultured cells as well as cell lines react in an oversensitive manner to the addition of TCFs.

scholarly journals Stimulation by cAMP of erythropoietin secretion by an established human renal carcinoma cell line

Blood ◽  
1987 ◽  
Vol 69 (4) ◽  
pp. 1053-1057 ◽  
Author(s):  
JB Sherwood ◽  
ER Burns ◽  
D Shouval
Keyword(s):  
Cell Line ◽  
Renal Carcinoma ◽  
Carcinoma Cell ◽  
Phosphodiesterase Inhibitor ◽  
Adenosine Monophosphate ◽  
Carcinoma Cell Line ◽  
Biologically Active ◽  
Human Renal Carcinoma ◽  
Renal Carcinoma Cell Line

Abstract We used our recently reported stable, transformed human renal carcinoma cell line as a model system to study the role of 3′,5′-adenosine monophosphate (cAMP) in erythropoietin secretion. The erythropoietin produced by these cells is both biologically active and immunologically cross-reactive with purified native human hormone in our radioimmunoassay. Erythropoietin release by these renal carcinoma cells appears to be stimulated by cAMP as well as by the phosphodiesterase inhibitor 3-isobutyl-1-methyl-xanthine (MIX). The response to cAMP involves a rapid and enhanced release of hormone, which occurred within 30 minutes of exposure of the cells to the effector and continued for at least 4 hours. Intracellular erythropoietin was higher in the control cultures than in the cells treated with cAMP, suggesting that cAMP stimulates the release of a storage pool of hormone. The ability of cAMP and MIX to elicit the release of erythropoietin suggests that a cAMP-mediated mechanism is involved in the release of this hormone.

scholarly journals Cardiac myofibroblasts isolated from the site of myocardial infarction express endothelin de novo

2003 ◽  
Vol 285 (3) ◽  
pp. H1132-H1139 ◽  
Author(s):  
Laxmansa C. Katwa
Keyword(s):  
Myocardial Infarction ◽  
Tissue Repair ◽  
Type I Collagen ◽  
Cultured Cells ◽  
De Novo ◽  
Culture Media ◽  
Myocardial Infarct ◽  
Biologically Active ◽  
Myocardial Infarct Size ◽  
Type I

Recently it was demonstrated that treatment with a nonselective endothelin (ET) receptor antagonist significantly reduces myocardial infarct size, which suggests a major role for ET in tissue repair following myocardial infarction (MI). Tissue repair and remodeling found at the site of MI are mainly attributed to myofibroblasts (myoFbs), which are phenotypically transformed fibroblasts that express α-smooth muscle actin. It is unclear whether myoFbs generate ET peptides and consequentially regulate pathophysiological functions de novo through expression of the ET-1 precursor (prepro-ET-1), ET-converting enzyme-1 (ECE-1), a metalloprotease that is required to convert Big ET-1 to ET-1 and ET receptors. To address these intriguing questions, we used cultured myoFbs isolated from 4-wk-old MI scar tissue. In cultured cells, we found: 1) expression of mRNA for ET precursor gene ( ppET1), ECE-1, and ETA and ETB receptors by semiquantitative RT-PCR; 2) phosphoramidon-sensitive ECE-1 activity, which converts Big ET-1 to biologically active peptide ET-1; 3) expression of ETA and ETB receptors; 4) elaboration of Big ET-1 and ET-1 peptides in myoFb culture media; and 5) upregulation of type I collagen gene expression and synthesis by ET, which was blocked by bosentan (a nonselective ETA- and ETB receptor blocker). These studies clearly indicated that myoFbs express and generate ET-1 and receptor-mediated modulation of type I collagen expression by ET-1. Locally generated ET-1 may contribute to tissue repair of the infarcted heart in an autocrine/paracrine manner.

scholarly journals Effects of dibutyryl adenosine 3',5'-cyclic monophosphate on erythropoietin production in human renal carcinoma cell cultures

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 714-717 ◽  
Author(s):  
M Hagiwara ◽  
SM Pincus ◽  
IL Chen ◽  
BS Beckman ◽  
JW Fisher
Keyword(s):  
Cell Cultures ◽  
Renal Carcinoma ◽  
Cultured Cells ◽  
Carcinoma Cell ◽  
Athymic Nude Mice ◽  
Dome Formation ◽  
Cyclic Monophosphate ◽  
Erythropoietin Production ◽  
Primary Monolayer ◽  
Human Renal Carcinoma

Abstract A human renal carcinoma from a patient with erythrocytosis, serially transplanted into athymic nude mice, was grown in primary monolayer cell cultures. After reaching confluency, the cultured cells formed multicellular hemicysts (domes), which became more abundant as the cultures approached saturation density. Erythropoietin (Ep) production by this renal carcinoma in culture was only slightly increased at the time of semiconfluency but showed a marked increase after the cultures reached confluency, in parallel with dome formation. Dibutyryl adenosine 3′,5′-cyclic monophosphate significantly (P less than .01) stimulated Ep production and dome formation in the semiconfluent and confluent cultures of the renal carcinoma.

scholarly journals Effects of dibutyryl adenosine 3',5'-cyclic monophosphate on erythropoietin production in human renal carcinoma cell cultures

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 714-717
Author(s):  
M Hagiwara ◽  
SM Pincus ◽  
IL Chen ◽  
BS Beckman ◽  
JW Fisher
Keyword(s):  
Cell Cultures ◽  
Renal Carcinoma ◽  
Cultured Cells ◽  
Carcinoma Cell ◽  
Athymic Nude Mice ◽  
Dome Formation ◽  
Cyclic Monophosphate ◽  
Erythropoietin Production ◽  
Primary Monolayer ◽  
Human Renal Carcinoma

A human renal carcinoma from a patient with erythrocytosis, serially transplanted into athymic nude mice, was grown in primary monolayer cell cultures. After reaching confluency, the cultured cells formed multicellular hemicysts (domes), which became more abundant as the cultures approached saturation density. Erythropoietin (Ep) production by this renal carcinoma in culture was only slightly increased at the time of semiconfluency but showed a marked increase after the cultures reached confluency, in parallel with dome formation. Dibutyryl adenosine 3′,5′-cyclic monophosphate significantly (P less than .01) stimulated Ep production and dome formation in the semiconfluent and confluent cultures of the renal carcinoma.

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