Evidence for the presence of viable endothelial cells in cultures derived from dissociated rat brain

Summary The morphology and histochemistry of dissociated newborn rat brain was studied in tissue culture. Direct microscopy of developing cells, electron microscopy and the alkaline phosphatase activity were used to identify the capillary endothelial cells.Acknowledgments. We thank Dr J. Gripenberg for technical assistance. This research was supported by Finnish Cultural Foundation and carried out during the tenure of a fellowship provided for F. J. from the Finnish-Hungarian Cultural Exchange Program.     

A study of irrigation fluids for neurosurgery on brain primary cell cultures

Summary Primary cell cultures from newborn rat brain hemispheres were exposed to different irrigation fluids used in neurosurgery. The cells died after incubation for 5 min with hydrogen peroxide, and the number of cells was drastically decreased after 10 sec of incubation. They shrank after incubation in Elliott's artificial cerebrospinal fluid for 3 h, but the viability as determined by trypan blue exclusion test was not affected. Physiological sodium chloride, Ringer's solution and the culture medium 199 with Hank's salt had no noticeable effect on the viability or morphology of the cells.Acknowledgments. This study was supported by grants from the Medical Faculty of Göteborg and from Statens Naturvetenskapliga Forskningsråd. We want to thank Dr Åke Sellström and Dr Hans-Arne Hansson for valuable discussions.     

Monoamine-containing neurones in cultures of rat brain stem

Zusammenfassung Monoaminhaltige Neurone konnten mit Hilfe der Fluoreszenz-Mikroskopie in Hirnstammgewebe, welches während mehrerer Tage in vitro gezüchtet wurde, nachgewiesen werden. Reserpin und Natriumborhydrid bewirkten eine deutliche Abnahme dieser Fluoreszenz. Diese Untersuchungen weisen darauf hin, dass Neurone, welche während mehrerer Tage in vitro gezüchtet wurden, die Fähigkeit haben, Monoamine zu speichern und/oder zu synthetisieren.     

Simplifications to substrate preparation for the cultivation of dissociated nerve cells

Résumé Nous avons simplifié la méthode de préparation du substrat «collagène» pour la culture de cellules nerveuses dissociées. On obtient une fixation et une différenciation rapide des cellules sur ce substrat. Les cultures conviennent pour des études morphologiques, biochimiques et électrophysiologiques.

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Chargée de Recherche au CNRS.

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Acknowledgments. The technical assistance of Mrs.Dorea van Weele andMrs. Marie-France Knoetgen is gratefully acknowledged.     

Brain chopping: A new method for the rapid removal of newborn rat brain

Summary A new multi-bladed air-driven guillotine is described. This device removes newborn rat heads and slices them into several thin wafers, which facilitates freezing and permits easy regional dissection.Supp. in part by U. S. Public Health Service Grant. AA-01391.     

A new intracellular medium for prolonged viability of noradrenergic storage vesicles from rat brain

Summary The biochemical lifetime of noradrenergic storage vesicles from rat brain is greatly extended in a medium based upon the membrane impermeable anion D-tartrate. Examination of the MgATP-dependent accumulation of (–)-³H-norepinephrine suggests that this medium may more closely approximate the intracellular environment of the storage vesicle, and may better support electrochemical gradients existing across the vesicle membrane.Acknowledgments. This work was supported by grant NS 15374 (NIH) of the United States Public Heath Service.     

A new synthesis of benzoyl phosphate: A substrate for acyl phosphatase assay

Summary A new method for the synthesis of benzoyl phosphate was reported. The advantages are: 1. more rapid procedure; 2. lower cost; 3. higher yield.This work was supported by grants from the Italian Consiglio Nazionale delle Ricerche and from the Ministero della Pubblica Istruzione.     

Steady-state distribution of lithium during cultivation of dissociated brain cells

Summary The formation of a steady-state intracellular lithium level was studied in the course of cultivation of dissociated nerve cell cultures obtained from chick embryonic brains. When lithium was given at a concentration of 2 mM, in the nutrient medium, at day 5, a steady state intracellular lithium content was achieved after about 30 min of incubation and it did not change significantly during the time of cultivation up to the 13th day in vitro.Acknowledgment. We thank Dr Latzkovits for useful advice and helping in methodology.     

Complete replacement of serum in primary cultures of chick embryo brain cells by growth-promoting alpha-globulin

Summary Growth-promoting alpha-globulin (GPAG), a specific serum protein complex which induces mitotic activity in continually replicating metazoan cells in vitro, was shown in this study to support growth of astrocytes and mesenchymal cells as well as process formation of nerve cells isolated from cerebral hemispheres of chick embryos.     

Complete replacement of serum in primary cultures of chick embryo brain cells by growth-promoting alpha-globulin

Growth-promoting alpha-globulin (GPAG), a specific serum protein complex which induces mitotic activity in continually replicating metazoan cells in vitro, was shown in this study to support growth of astrocytes and mesenchymal cells as well as process formation of nerve cells isolated from cerebral hemispheres of chick embryos.     

Participation of microtubules and microfilaments in the transcellular biliary secretion of immunoglobulin A in primary cultures of rat hepatocytes

Summary The biliary secretion of immunoglobulin A (IgA) in primary hepatocyte cultures was investigated by means of immunofluorescence. The characteristic accumulation of IgA in visible bile canaliculi was found to be strongly inhibited by vinblastine and colchicine or by cytochalasin B, but its surface binding was not.The author thanks Prof. Dr D. Mecke for his support and encouragement. The excellent technical assistance of Mrs M. Behringer and Mrs A. Schneck is gratefully acknowledged. The author is grateful to the BIOTEST Serum Institut (Frankfurt) for the kind gift of human IgA. This work was supported by the Deutsche Forschungsgemeinschaft.     

A simple procedure for removal of intra- and extra-tumoral macrophages from primary tumor cultures

Summary Improved growth of tumor cells in primary cell suspensions isolated from solid tumors can be achieved by addition of an antifungal agent, mycostatin, to the culture medium. Mycostatin inhibits the cytotoxicity of macrophages and also quickly detaches phagocytes without any apparent retardation of the growth of tumor cells. This method is particularly useful for obtaining cultures of lung tumor cells or cells from lung metastases of other tumors.This research was supported in part by grants CA-22839 and CA-28482 awarded by the National Cancer Institute, DHEW, USA. This is publication No. 406 from the Department of Basic and Clinical Immunology and Microbiology, Medical University of South Carolina.We thank Charles L. Smith for editorial assistance. Address correspondence to Y. Y., Department of Basic and Clinical Immunology and Microbiology, Medical University of South Carolina, 171 Ashley Avenue, Charleston, SC 29403, USA.     

Histochemical study of dissociated nerve cells from embryonic chick cerebral hemispheres in flask cultures

Zusammenfassung Die Differenzierung von dissoziierten Grosshirnneuronen in Plastikflaschenkulturen wurde mit histochemischen Methoden, Nisslfärbung, Silberimprägnation der Nervenfasern und Acetylcholinesteraseaktivität untersucht.

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with the technical assistance ofM. F. Knoetgen

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Chargée de Recherche au C.N.R.S.     

Brain chopping: a new method for the rapid removal of newborn rat brain.

A new multi-bladed air-driven guillotine is described. This device removes newborn rat heads and slices them into several thin wafers, which facilitates freezing and permits easy regional dissection.     

Effect of glucocorticoids on the appearance of gamma-glutamyl transpeptidase activity in primary cultures of adult rat hepatocytes

Primary cultures of adult rat liver parenchymal cells showed a progressive rise of gamma-glutamyl transpeptidase (GGT) activity (E.C. 2.3.2.2) after the first 5 days of culture. The presence of dexamethasone and other synthetic glucocorticoids in the culture medium partially prevented this increase.