Cell surface heparan sulphate and adhesive property of sublines of rat ascites hepatoma AH7974

1988 ◽  
Vol 90 (4) ◽  
pp. 683-689 ◽  
Author(s):  
A. Kimura ◽  
T. Kawaguchi ◽  
T. Ono ◽  
A. Sakuma ◽  
Y. Yokoya ◽  
...  
Keyword(s):  
Cell Surface ◽  
Culture Medium ◽  
Heparan Sulphate ◽  
Soluble Form ◽  
Foetal Calf Serum ◽  
Serum Free Medium ◽  
Free Medium ◽  
Ascites Hepatoma ◽  
Serum Free ◽  
Rat Ascites Hepatoma

Two variants (74AD and 74FL) established from rat ascites hepatoma AH7974 were examined for the production of glycosaminoglycans in culture. There was no difference between the adhesive (74AD) and the floating (74FL) variants in quantity of glycosaminoglycans produced by their cultivation in minimum essential medium supplemented with 10% foetal calf serum. However, they were distinctly different in the distribution patterns of heparan sulphate. In 74FL, about 70% of total heparan sulphate was found in the culture medium in soluble form, whereas in 74AD, only 7% was found in the medium and the rest was in the cell-substratum complex. In a serum-free medium, 74AD cells grew without adhering to the substratum. After cultivation, more than 90% of total heparan sulphate was found in the cell-associated fractions and the rest in the substratum fractions. No heparan sulphate was detected in the culture medium. On the other hand, 74FL cells released heparan sulphate to the serum-free medium as much as to the serum-containing medium. The increase in amount of heparan sulphate in the culture medium of 74FL cells was supposed to be caused by failure of the cells to deposit heparan sulphate at the cell surface and not caused by increased production. Cell-substratum adhesion mechanisms involving cell surface heparan sulphate (heparan sulphate proteoglycan) and some serum intermediate(s) are discussed for 74AD cells.

scholarly journals Stimulierung von Kulturen embryonaler Rattenzellen durch eine Proteinfraktion aus fötalem Kälberserum / Stimulation of Embryonic Rat Cells in Culture by a Protein Fraction Isolated from Fetal Calf Serum

1971 ◽  
Vol 26 (10) ◽  
pp. 1045-1048 ◽  
Author(s):  
Dieter F. Hülser ◽  
Werner Frank
Keyword(s):  
Cell Cycle ◽  
Culture Medium ◽  
Fetal Calf Serum ◽  
Surface Membrane ◽  
Calf Serum ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Membrane Potential Difference ◽  
Stimulation Of

Normal embryonic rat cells incubated in serum-free medium accumulate in G1-phase of the cell cycle. On addition of a growth-stimulating protein isolated from fetal calf serum they are triggered to proceed through the cycle, and they resume DNA-synthesis 15 to 20 hours later. In this paper it is demonstrated that the surface membrane potential difference (PD) decreases immediately after changing serum-free medium against culture medium containing either calf serum or the isolated serum protein; the original PD is restored 2 to 3 hours later. Serumprotein without growthstimulating activity does not affect the PD.A permanent rat cell line which grows independently of serum also has been tested. The PD of these cells is not significantly influenced by calf serum.

scholarly journals Investigating the Interaction of Nanodiamonds with Human Serum Albumin and Induced Cytotoxicity

2019 ◽  
Vol 2019 ◽  
pp. 1-9 ◽  
Author(s):  
Miaomiao Chen ◽  
Xiaoshuang Zuo ◽  
Qinqin Xu ◽  
Rong Wang ◽  
Suhua Fan ◽  
...  
Keyword(s):  
Cell Culture ◽  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Biomedical Applications ◽  
Culture Medium ◽  
Cell Culture Medium ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free

Nanodiamonds (NDs) have been recognized as emerging carbon-based delivery vehicles due to their biocompatibility. NDs were reported to be nontoxic and suited for biomedical applications in the complete cell culture medium. However, in this study, the cytotoxicity of NDs in serum-free medium was studied and indicated that serum proteins in cell culture medium played significant effect on the toxicity of NDs. Therefore, the interaction mechanism between NDs and a serum protein (human serum albumin, HSA) was first investigated by fluorescence quenching technique and circular dichroism (CD) spectrometry. The results suggest that HSA strongly bonds on the NDs surface to form “protein corona,” which not only prevents the aggregation of NDs and improves its stability but also inhibits the cytotoxicity of NDs. In serum-free medium, NDs exhibited obvious toxicity toward the human lung epithelial cell line (BEAS-2B) and showed concentration-dependent cytotoxicity. In the presence of bovine serum albumin (BSA), which shares structural homology and similar properties with HSA, toxicity of NDs was apparently inhibited. Therefore, the interaction between serum protein and NDs should be considered in the understanding of the biological effects of NDs exposure in biomedical applications.

scholarly journals Changes in sterol biosynthesis accompanying cessation of glial cell growth in serum-free medium

1980 ◽  
Vol 192 (2) ◽  
pp. 709-717 ◽  
Author(s):  
William A. Maltese ◽  
Beverly A. Reitz ◽  
Joseph J. Volpe
Keyword(s):  
Fatty Acid ◽  
Reductase Activity ◽  
Calf Serum ◽  
Sterol Synthesis ◽  
Foetal Calf Serum ◽  
Quiescent State ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Coa Reductase

C-6 glioma cells, grown in medium supplemented with 5% delipidated foetal calf serum, were induced to enter a quiescent state by removing serum from the medium. Within 24h there was a 75–80% decline in the rate of incorporation of [14C]acetate or 3H2O into digitonin-precipitable sterols. Experiments with [3H]mevalonolactone as a labelled sterol precursor suggested that the decline in sterol synthesis was regulated primarily at a point in the pathway before the formation of mevalonate. The specific activities of 3-hydroxy-3-methylglutaryl-CoA synthase and 3-hydroxy-3-methylglutaryl-CoA reductase decreased sharply in conjunction with the decline in sterol synthesis in the serum-free cultures; however, the activity of acetoacetyl-CoA thiolase was altered only slightly. The magnitude of the initial decline in reductase activity was not affected when 50-mm-NaF was included in the preincubation and assay buffers to prevent activation of physiologically inactive enzyme. However, after 6h of serum deprivation the decline in 3-hydroxy-3-methylglutaryl-CoA reductase activity was due to a decrease in the amount of latent activity. The sterol concentration in C-6 cells was unchanged after 24h in serum-free medium, although a 20% decrease in the sterol/fatty acid molar ratio occurred as a result of a small increase in the fatty-acid concentration. Incorporation of 3H2O into fatty acids was inhibited in the serum-deprived glial cells; however, this inhibition developed more slowly and was not as pronounced as the diminution in sterol synthesis. The results suggest that in C-6 glia, which resemble the glial stem cells of the developing brain, the decreased demand for membrane sterols in the quiescent state results in a decline in sterol synthesis, mediated primarily through co-ordinate changes in the activities of 3-hydroxy-3-methylglutaryl-CoA synthase and 3-hydroxy-3-methylglutaryl-CoA reductase.

Fasciola hepatica in vitro: increased susceptibility to fasciolicides in a defined serum-free medium

Parasitology ◽  
1987 ◽  
Vol 95 (1) ◽  
pp. 165-171 ◽  
Author(s):  
D. C. Jenkins ◽  
P. Topley ◽  
E. B. Rapson
Keyword(s):  
Culture Medium ◽  
Fasciola Hepatica ◽  
Liver Microsomes ◽  
Metabolic Activation ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Highly Active ◽  
Mammalian Liver

SUMMARYThe cidal properties of some phenolic, halogenated diphenyl, salicylanilide, benzimidazole and diaminophenoxyalkane anthelmintics, against 6-week-old worms of Fasciola hepatica were assessed in vitro. In a conventional fluke culture medium containing RPMI 1640, supplemented with serum with or without rabbit erythrocytes or pink-ghosts, only the halogenated diphenyl and salicylanilide compounds showed activity at concentrations equal to or less than 100 μm. However, when basal, serum and cell-free RPMI 1640 was used, all compounds other than diamphenethide were highly active, their minimum lethal concentrations being some 25–125 times lower under these conditions. The inclusion of rabbit liver microsomes in the basal culture medium resulted in diamphenethide exhibiting cidal activity equivalent to that seen when its free-amine active metabolite was assayed. The possibility that the activity of many of these compounds was masked in vitro because of their serum binding properties is discussed. Recommendations are made that in vitro screens for new fasciolicides should be carried out in serum-free medium and that additional replicates containing mammalian liver microsomes and liver cytosolic extracts be included as means for the metabolic activation of certain otherwise undetectable prodrugs.

scholarly journals Mitogenic agents induce redistribution of transferrin receptors from internal pools to the cell surface

1986 ◽  
Vol 238 (3) ◽  
pp. 721-728 ◽  
Author(s):  
D McV Ward ◽  
J Kaplan
Keyword(s):  
Growth Factor ◽  
Cell Surface ◽  
Transferrin Receptor ◽  
Transferrin Receptors ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Surface Receptor ◽  
Receptor Number ◽  
Receptor Distribution

Incubation of serum-growth HeLa cells in serum-free medium causes a rapid (t1/2 3 min) 30-60% decrease in the binding of 125I-diferric transferrin to the cell surface. Addition of fetal bovine serum to cells in serum-free medium results in a rapid (t1/2 3 min) and concentration-dependent increase in binding activity. The loss or gain in ligand binding is a result of changes in surface receptor number rather than an alteration in ligand-receptor affinity. A variety of hormones (insulin, insulin-like growth factor, interleukin 1 and platelet-derived factor) were found to mimic the effect of serum on receptor number. The alteration in surface receptor number was found to be calcium-dependent. Changes in surface receptor number were independent of either receptor biosynthetic rate or the absolute cellular content of receptors. The effect of insulin or serum on Hela cell transferrin receptor distribution was unaffected by the presence of transferrin, demonstrating that receptor distribution in this cell type is ligand-independent. The ability of serum or insulin to modify surface transferrin receptor number was also observed in mouse L-cells, human skin fibroblasts, and J774 macrophage tumour cells. However, transferrin receptors on K562 and Epstein-Barr virus-transformed human lymphoblasts were unaltered by these agents. The quantities of receptors whose distribution is predominantly on the surface (i.e. epidermal growth factor or low density lipoprotein receptor) were unaltered by addition of the mitogenic agents. These results extend our previous studies [H.S. Wiley & J. Kaplan (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 7456-7460] demonstrating that mitogenic agents can induce redistribution of receptor pools in selected cell types.

scholarly journals Extraction of Cholesterol with Methyl-β-Cyclodextrin Perturbs Formation of Clathrin-coated Endocytic Vesicles

1999 ◽  
Vol 10 (4) ◽  
pp. 961-974 ◽  
Author(s):  
Siv Kjersti Rodal ◽  
Grethe Skretting ◽  
Øystein Garred ◽  
Frederik Vilhardt ◽  
Bo van Deurs ◽  
...  
Keyword(s):  
Immunogold Labeling ◽  
Water Soluble ◽  
Soluble Form ◽  
Transferrin Receptors ◽  
Serum Free Medium ◽  
Free Medium ◽  
Coated Pits ◽  
Serum Free ◽  
Electron Microscopical ◽  
Endocytic Vesicles

The importance of cholesterol for endocytosis has been investigated in HEp-2 and other cell lines by using methyl-β-cyclodextrin (MβCD) to selectively extract cholesterol from the plasma membrane. MβCD treatment strongly inhibited endocytosis of transferrin and EGF, whereas endocytosis of ricin was less affected. The inhibition of transferrin endocytosis was completely reversible. On removal of MβCD it was restored by continued incubation of the cells even in serum-free medium. The recovery in serum-free medium was inhibited by addition of lovastatin, which prevents cholesterol synthesis, but endocytosis recovered when a water-soluble form of cholesterol was added together with lovastatin. Electron microscopical studies of MβCD-treated HEp-2 cells revealed that typical invaginated caveolae were no longer present. Moreover, the invagination of clathrin-coated pits was strongly inhibited, resulting in accumulation of shallow coated pits. Quantitative immunogold labeling showed that transferrin receptors were concentrated in coated pits to the same degree (approximately sevenfold) after MβCD treatment as in control cells. Our results therefore indicate that although clathrin-independent (and caveolae-independent) endocytosis still operates after removal of cholesterol, cholesterol is essential for the formation of clathrin-coated endocytic vesicles.

Evidence for a single glucocorticoid regulated pool of adrenocorticotropin in sheep anterior pituitary

1995 ◽  
Vol 268 (1) ◽  
pp. E85-E91
Author(s):  
R. J. Kemppainen ◽  
T. P. Clark
Keyword(s):  
Anterior Pituitary ◽  
Adrenocorticotropic Hormone ◽  
Culture Medium ◽  
Pituitary Cells ◽  
Serum Free Medium ◽  
Free Medium ◽  
Isolated Cells ◽  
Serum Free ◽  
Fractional Release ◽  
Acth Release

The goal of this study was to determine whether separate glucocorticoid-sensitive releasable pools of adrenocorticotropic hormone (ACTH) could be distinguished in sheep anterior pituitary cells. Isolated cells were cultured in serum-free medium containing 0–10 nM cortisol (F) for 7–11 days to determine whether variation in the glucocorticoid environment selectively affected ACTH release stimulated by corticotropin-releasing hormone (CRH) or arginine vasopressin (AVP). Secretion was studied using a microperifusion system. The results indicated that while the concentration of F in the medium bathing the cells profoundly influenced the magnitude of ACTH released in response to either peptide, the fractional release of total ACTH was unchanged. F concentration in culture medium similarly did not alter the negative-feedback effectiveness of a larger dose of F applied to cells 45 min before treatment with CRH or AVP. These results support the existence of a single glucocorticoid-sensitive pool of ACTH in corticotrophs.

scholarly journals THE REQUIREMENT FOR HYDROCORTISONE IN ANTIBODY-FORMING TISSUE CULTIVATED IN SERUM-FREE MEDIUM

1964 ◽  
Vol 119 (6) ◽  
pp. 1027-1049 ◽  
Author(s):  
Charles T. Ambrose
Keyword(s):  
Culture Medium ◽  
Essential Amino Acids ◽  
Secondary Response ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Secondary Antibody ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free

It was previously reported from this laboratory that the secondary antibody response can regularly be elicited in vitro from fragments of rabbit lymph node node cultured in Eagle's medium supplemented with normal rabbit serum. Evidence is now presented that physiological levels of hydrocortisone (0.01 to 1.0 µM) can substitute for serum in the culture medium. However, with the omission of serum, serine (0.1 mM) must be included among Eagle's "essential" amino acids for consistent optimal antibody production. In some experiments the addition of insulin (0.5 unit/ml) and vitamin B12 (0.5 µg/ml) has further enhanced the secondary response in this serum-free medium.

scholarly journals Development of a Biodegradable Microcarrier for the Cultivation of Human Adipose Stem Cells (hASCs) with a Defined Xeno- and Serum-Free Medium

2021 ◽  
Vol 11 (3) ◽  
pp. 925
Author(s):  
Francesco Muoio ◽  
Stefano Panella ◽  
Matias Lindner ◽  
Valentin Jossen ◽  
Yves Harder ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Culture Medium ◽  
Free Cell ◽  
Adipose Stem Cells ◽  
Cell Culture Medium ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Human Adipose Stem Cells

Stirred single-use bioreactors in combination with microcarriers (MCs) have established themselves as a technology that has the potential to meet the demands of current and future cell therapeutic markets. However, most of the published processes have been performed using fetal bovine serum (FBS) containing cell culture medium and non-biocompatible MCs. This approach has two significant drawbacks: firstly, the inevitable potential risks associated with the use of FBS for clinical applications; secondly, non-biocompatible MCs have to be removed from the cell suspension before implantation, requiring a step that causes loss of viable cells and adds further costs and complications. This study aimed to develop a new platform based on a chemically defined xeno- and serum-free cell culture medium and biodegradable MC that can support the growth of human adipose stem cells (hASCs) while still preserving their undifferentiated status. A specific combination of components and manufacturing parameters resulted in a MC prototype, called “BR44”, which delivered the desired functionality. MC BR44 allows the hASCs to stick to its surface and grow in a chemically defined xeno- and serum-free medium (UrSuppe). Although the cells’ expansion rate was not as high as with a commercial non-biodegradable standard MC, those cultured on BR44 maintained a better undifferentiated status in both static and dynamic conditions than those cultured on traditional 2D surfaces.

Production of somatomedin-like activity by human adult tumor-derived, transformed, and normal cell cultures and by cultured rat hepatocytes: effects of culture conditions and of epidermal growth factor (urogastrone)

1984 ◽  
Vol 62 (12) ◽  
pp. 1343-1350 ◽  
Author(s):  
Paul R. Atkison ◽  
L. Joseph Hayden ◽  
R. Marvin Bala ◽  
Morley D. Hollenberg
Keyword(s):  
Human Skin ◽  
Cell Cultures ◽  
Culture Medium ◽  
Rat Hepatocytes ◽  
Serum Free Medium ◽  
Free Medium ◽  
Kb Cells ◽  
Serum Free ◽  
Epidermal Growth ◽  
Adult Human Skin

We have measured the production of a basic-somatomedin-like activity (SLA) by a variety of human tumor-derived, transformed, and normal postnatal cell cultures; and we have compared the production of SLA by these cell types with the production of SLA by adult rat hepatocytes cultured in serum-free medium. Cells derived from a human epidermoid carcinoma (KB), a pancreatic carcinoma (Panc-1), a Simian virus 40 transformed adult human skin-derived cell line (SV40 fibroblasts), and a normal adult human skin-derived fibroblast line released SLA when cultured in a serum-free growth medium. No SLA was recovered from the culture medium of human choriocarcinoma-derived cells (BeWo) or of a human lymphoblastoid cell line (IM-9). The production of SLA by rat hepatocytes cultured in serum-free medium appeared to exceed the production of SLA by the other cell cultures. In cultures of KB cells, SV40 fibroblasts, and rat hepatocytes, the production of SLA depended on the frequency with which the growth medium was renewed; in general, the highest rates of SLA production were observed when the medium was renewed every 48–72 h. The presence of mouse epidermal growth factor (urogastrone) (EGF-URO) in the serum-free culture medium stimulated the production of SLA by KB cells and by rat hepatocytes, but did not increase SLA production by normal or by SV40-transfonned human skin-derived fibroblasts. We conclude that tumor-derived cells are capable of producing somatomedin-like activity and that the production of SLA by such cells can be subject to controls (nutrient availability, EGF-URO stimulation) that regulate SLA production, either by normal adult tissues, like liver, or by a variety of normal embryonic tissues.

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