Production Of CSF Research Articles

Induction of growth alterations in factor-dependent hematopoietic progenitor cell lines by cocultivation with irradiated bone marrow stromal cell lines

We studied the production of hemopoietins by x-irradiated plateau-phase cultures of cloned marrow stromal cell lines derived from C3H/HeJ marrow, termed D2XRII and clone 11. The production of CSF in agar overlay of control or 10,000 rad irradiated stromal cultures was quantitated by induction of colonies in: overlaid fresh marrow, IL-3-dependent cell line 32D cl 3, or GM-CSF/IL-3-dependent cell lines FDCP-1 or bg/bg cl 1. Conditioned media were tested for CSF by bioassay using fresh marrow cells, for M-CSF (CSF-1) by RIA, and for IL-3 and GM-CSF by microwell proliferation assay with 32D cl 3 and FDCP-1 cells, respectively. X-irradiation to doses that decreased CSF-1 to 40% of control levels (greater than 5,000 rad) resulted in a 30-fold increase in growth of FDCP-1 or bg/bg cl 1 cells in liquid co-culture or agar culture overlay with no detectable growth of 32D cl 3. The frequency of subculture of nonautocrine, factor independent (FI) variant clonal lines of FDCP-1 or bg/bg cl 1 cells was increased over 1000-fold by 15 weeks cocultivation with irradiated stromal cell cultures. FI subclonal lines formed tumors in syngeneic mice and contained no detectable poly A messenger RNA for GM-CSF or IL-3, and no elevation in c-myc, c-abl, c-src, or erb-B onc gene-specific messenger RNA compared to parent factor-dependent lines. These data indicate that x-irradiated plateau phase marrow stromal cells produce increased levels of cell contact-mediated biologically active hemopoietin(s) other than M-CSF, GM-CSF, or IL-3 and induce nonautocrine factor-independent malignant cell lines in vitro.

Induction of Growth Alterations in Factor-Dependent Hematopoietic Progenitor Cell Lines by Cocultivation With Irradiated Bone Marrow Stromal Cell Lines

Abstract We studied the production of hemopoietins by x-irradiated plateau-phase cultures of cloned marrow stromal cell lines derived from C3H/HeJ marrow, termed D2XRII and clone 11. The production of CSF in agar overlay of control or 10,000 rad irradiated stromal cultures was quantitated by induction of colonies in: overlaid fresh marrow, IL-3- dependent cell line 32D cl 3, or GM-CSF/IL-3-dependent cell lines FDCP- 1 or bg/bg cl 1. Conditioned media were tested for CSF by bioassay using fresh marrow cells, for M-CSF (CSF-1) by RIA, and for IL-3 and GM- CSF by microwell proliferation assay with 32D cl 3 and FDCP-1 cells, respectively. X-irradiation to doses that decreased CSF-1 to 40% of control levels (greater than 5,000 rad) resulted in a 30-fold increase in growth of FDCP-1 or bg/bg cl 1 cells in liquid co-culture or agar culture overlay with no detectable growth of 32D cl 3. The frequency of subculture of nonautocrine, factor independent (FI) variant clonal lines of FDCP-1 or bg/bg cl 1 cells was increased over 1000-fold by 15 weeks cocultivation with irradiated stromal cell cultures. FI subclonal lines formed tumors in syngeneic mice and contained no detectable poly A messenger RNA for GM-CSF or IL-3, and no elevation in c-myc, c-abl, c- src, or erb-B onc gene-specific messenger RNA compared to parent factor- dependent lines. These data indicate that x-irradiated plateau phase marrow stromal cells produce increased levels of cell contact-mediated biologically active hemopoietin(s) other than M-CSF, GM-CSF, or IL-3 and induce nonautocrine factor-independent malignant cell lines in vitro.

Differentiation and production of colony-stimulating factor induced by immunostimulants in a leukemia cell line.

Immunostimulants from various microorganisms were tested on a myeloid leukemia cell line (M1) for the ability to induce production of CSF and to cause differentiation of these cells. Based on their activities, the compounds were divided into two general classes: those inducing extensive cellular differentiation and those devoid of this effect. The stimulants which were active in this regard always produced large quantities of CSF, whereas those devoid of a differentiating effect did not cause CSF production. Even the potent stimulants had no effect on the D subline, in which the differentiation was not inducible.

A Case of Lung Cancer Associated with Granulocytosis and Production of CSF from the Tumor

A Case of Lung Cancer Associated with Granulocytosis and Production of CSF from the Tumor

Changes in enzyme activities of the choroid plexus and brain tissue following bilateral carotid ligation in correlation with the formation of cerebrospinal fluid.

The activity of four enzymes which are thought to be correlated to the production of CSF, such as acid and alkaline phosphatase, carbonic anhydrase, and Na-K ATPase, were determined biochemically in the choroid plexus and the frontal lobe brain tissue in adult rabbits. The activities of these enzymes, except Na-K ATPase, were transiently enhanced in the choroid plexus after bilateral ligation of the common carotid arteries, although considerable decrease was found in the brain tissue. Since the above results had suggested that enhancement of secretory activity of the choroid plexus followed bilateral carotid ligation, the rate of CSF formation was measured by means of the ventriculocisternal perfusion method in the same species of animals before and after bilateral carotid ligation. However, no significant changes were observed in the rate of CSF formation until 2 hours after carotid ligation. After bilateral occlusion of the common carotid arteries, vertebral blood flow, measured electromagnetically, immediately increased to twice its normal rate. Also, resin casts made of the choroid plexus after bilateral carotid ligation, confirmed that there was an adequate blood supply to the entire choroid plexus through the vertebral-posterior choroidal artery system. Considering the dual source of CSF production, it is likely that the secretory activity of the choroid plexus not only is unaffected by a sudden change of cerebral blood flow but also may compensate for the reduced production of CSF in the brain for at least a temporary period. Therefore, in face of ischemic-anoxic changes in the brain this could lead to the maintenance of a normal rate of CSF production by a homeostatic control mechanism.

Transfer of I125-albumin from blood into brain and cerebrospinal fluid in newborn and juvenile rats.

Human fetuses, rabbit fetuses and newborn rats have a relatively high concentration of protein in cerebrospinal fluid (csf) as compared with the adult. The present study was undertaken in an attempt to evaluate whether this high protein concentration in newborn rats is caused by a high permeability of the blood-brain-csf barrier to protein or by a low production rate of csf. The results suggest that the high concentration protein in csf in newborn rats is due to a low rate of production of csf rather than to an increased transfer of protein across the blood-brain-csf barrier.

Mediated transport of chloride from blood into cerebrospinal fluid.

The kinetics of transport of chloride from blood into CSF have been studied in the adult cat in vivo for a wide range of concentrations of chloride in arterial plasma: 1. In studies employing isosmotic replacement of body chloride with isethionate by means of extracorporeal hemodialysis, the chloride content in cerebral cortex and corpus callosum and concentration in CSF were relatively resistant to change despite the fact that plasma chloride was varied over the range 120.5 to 8.1 mM. 2. During hemodialysis the ratio of the concentration of chloride in CSF (or content in cerebral cortex) versus the concentration of chloride in arterial plasma was maintained for at least 1 or 2 h at as much as 10 times the ratios present in normal animals. 3. In studies with 36Cl, the rate of entry of chloride from blood into CSF followed Michaelis-Menten kinetics with the Vmax=0.55 μmole/ ml/min and with the Km < 60 mM and possibly < 8.1 mM. On the other hand the rate of entry of chloride from blood into cerebral cortex and corpus callosum was much slower and followed simple diffusion kinetics. 4. Evidence is presented indicating that chloride is transported from blood to CSP at the choroid plexus as well as at other sites. 5. On the basis of the rate of entry of chloride into CSF, the rate of bulk production of CSF in the adult cat was calculated to be 17.1 μl/min at normal plasma concentrations of chloride, a value in close agreement with previous estimates of the rate of production of CSF in the adult cat. 6. The relationships of the content of chloride in skeletal muscle and in liver to the concentration of chloride in plasma are presented, when the latter is varied over the range 120.5 to 8.1 mM.

Studies on the dynamics of the cerebrospinal fluid using radioactive isotopes. I. The formation and the absorption of cerebrospinal fluid.

Upon studying the relation between the production of CSF and the intracranial pressure in patients with various surgical brain diseases and the effect of intravenous injection of 50% glucose solution on the production, by following up the turnover of P-32 from blood to CSF, it was confirmed that CSF production was of course inactive in the cases without increased intracranial pressure, but in the cases with increased intracranial pressure too, the production was not always correlated with the rise of the intracranial pressure. The production was particularly active in the highpressure cases with advanced destruction of the blood-CSF barrier, as in the malignant glioma cases, but relatively inactive in the cases with equally high intracranial pressure due to obstruction of the aqueduct of Sylvius but apparently mild destruction of the blood-CSF barrier. Upon injection of 50% glucose solution, the production of CSF was inhibited to some extent in all cases, the inhibition being particularly notable in the cases with drastic destruction of the blood-CSF barrier.

PHYSIOLOGY OF THE CSF

SummaryA certain production of CSF occurs at the site of the choroid plexus. This CSF differs in composition from a pure plasma dia‐lysate and is thus probably formed by a secretory or other active process from the epithelium of the plexus. At the same time, however, certain substances, especially water, are in rapid exchange with the extracellular tissue of the brain and spinal cord.Reabsorption of the CSF occurs partly via arachnoidal villi and Pacchionian bodies to the cranial venous sinuses, partly via intrathecal capillaries and veins and finally to some extent by way of lymphatics via the perineural spaces.The protein in the CSF probably consists of serum protein, that has entered the CSF space. Protein is normally not formed within the central nervous system but may be under certain pathologic conditions. Normally, the serum protein enters the CSF space in two ways: pardy via meningeal vessels above all of finely dispersed proteins especially albumin, and partly via the choroid plexus either by passage via intercellular leaks in the plexus epithelium or also through secretion or some other active process. The CSF formed by the choroid plexus is relatively poor in protein and has an electrophoretic pattern, that differs more from that of serum protein than does CSF fluid from other parts of the CSF space.The reabsorption of the CSF protein occurs mainly directly to the blood stream, either via arachnoidal villi to the cranial venous sinuses or via intrathecal capillaries and veins. Reabsorption of protein via lymphatics is probably insignificant.