The viability of cells grown or centrifuged in a new density gradient medium, Percoll(TM)

Pertoft, Håkan; Rubin, Kristofer; Kjellén, Lena; Laurent, Torvard C.; Klingeborn, B.

doi:10.1016/0014-4827(77)90311-1

Cited by 215 publications

(50 citation statements)

References 9 publications

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“…Our findings concerning phagoeytic killing (Table 2) are in agreement with this. In contrast, FicoU is not a completely benign agent and can adversely affect the metabolism of leucocytes (Pertoft et al, 1977). Percoll is also taken up less than Ficoll by phagoeytic cells (Pertoft et al, 1977).…”

Section: Assessment Of Intracellular Killing Of P Mirabilis After Tementioning

confidence: 99%

Evaluation of Intracellular Killing of Bacteria by Enriched Populations of Mouse Peritoneal Exudate Neutrophils

Hart

Spencer

McDonald

et al. 1985

Aust J Exp Biol Med

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Summary. Elicited, mouse peritoneal exudate cells were fractionated by centrifugation on discontinuous Percoll density gradients. TVo subpopulations of neutrophils, each of greater than 90% purity, were isolated at discontinuous density gradient interfaces different from the region of mononuclear ceU enrichment (i.e., 1-0694-1 0871 and 1-0872-1-1002 g/ml for neutrophils and less than 1-0694 g/ml for mononuclear cells). Peritoneal exudate cells were mixed with Proteus mirabitis in the presence of 1% normal mouse serum for 30 min. The mixtures were fractionated on gradients of Percoll diluted with a clacium-free medium. Populations of cells banding at densities greater than 1 -0693 g/ml were washed free of gradient material, and neutrophil suspensions containing intracellular bacteria and which were relatively free of extracellular bacteria were isolated. Less than 7% of the total bacteria present was extracellular. The continuing extracellular presence of a heat-labile component of normal mouse serum was essential for maximal intracellular kill of P. mirabitis by mouse peritoneal neutrophils.

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Section: Assessment Of Intracellular Killing Of P Mirabilis After Tementioning

confidence: 99%

Evaluation of Intracellular Killing of Bacteria by Enriched Populations of Mouse Peritoneal Exudate Neutrophils

Hart

Spencer

McDonald

et al. 1985

Aust J Exp Biol Med

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“…For culture, cryopreserved cells were quickly thawed at 37°C and washed with M199 medium containing 10% FBS. The mammary epithelial acini were separated by a preformed 45% Percoll density gradient (Pharmacia, Uppsala, Sweden) centrifugation at 20,000 × g for 1 hr as described [8,19]. Mammary cells recovered from the Percoll density fraction higher than 1.055 g/ml were resuspended with M199 containing 15% FBS, insulin (5 µg/ ml, Sigma, St. Louis, MO, U.S.A.), hydrocortisone (5 µg/ ml, Sigma), ovine prolactin (1 µg/ml, Sigma), gentamicin (50 µg/ml), and amphotericin B 1 (2.5 µg/ml) and plated at a concentration of 2.5 × 10 5 cells/0.5 ml onto collagen-coated plastic 24-well cell culture plates (Iwaki Grass Co., Chiba, Japan).…”

mentioning

confidence: 99%

Detection of Interleukin-1 and Interleukin-6 on Cryopreserved Bovine Mammary Epithelial Cells in Vitro.

Okada

Ito

Ohtsuka

et al. 1997

The Journal of Veterinary Medical Science

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ABSTRACT. This investigation was performed to determine whether primary cultures of mammary cells from lactating cows would sustain production of interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF), and express mRNA for cytokines interleukin (IL)-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, interferon (INF)-τ, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF) by the reverse transcription-polymerase chain reaction (RT-PCR) in vitro. Cryopreserved mammary epithelial cells collected from cows at 1 week post calving were plated in collagen-coated 24-well culture plates (250,000 cells/well). IL-1 and IL-6 productions were measured using a A375 cell growth inhibition assay and a 7TD1 hybridoma proliferation assay, respectively. Production of IL-1 was demonstrated in mammary epithelial cells cultured with unsupplemented medium, but was not produced by cells cultured in medium supplemented with fetal bovine serum. IL-6 production in the conditioned medium was continued at steady level until day 14, whereas IL-6-like bioactivity was not detected in medium alone. TNF-like activity was not detectable in any experiments. This study also demonstrated the expression of mRNA for multiple cytokines including IL-1α, IL-1β, IL-6, IL-10, TNF-α, and GM-CSF by RT-PCR in mammary cell cultures. The results indicate that bovine mammary epithelial cells of lactating cows produce IL-1 and IL-6 and have gene expression for multiple cytokines. This in vitro model will be useful to investigate the function and regulation of IL-1 and IL-6 in the lactating mammary gland. -KEY WORDS: bovine, cytokine, mammary.

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“…Density gradients of Percoll were selected for purification of the cell fractions because of the low toxicity of this medium (22,23). The method adopted was as follows: Waymouth's medium dry powder (amount for 1 liter) was dissolved in 333 ml water and filtered through a 0.45-,um filter (Nalgene, Industrial Div.…”

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confidence: 99%

Modulation of a Glycoprotein Recognition System on Rat Hepatic Endothelial Cells by Glucose and Diabetes Mellitus

Summerfield¹,

Vergalla²,

Jones³

1982

J. Clin. Invest.

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A B S T R A C T The cellular location and carbohydrate specificities of a glycoprotein recognition system on rat hepatic sinusoidal cells have been determined. Purified preparations of endothelial, Kupffer, and parenchymal cells were prepared by collagenase liver perfusion, centrifugation on Percoll gradients, and centrifugal elutriation. '251-labeled agalactoorosomucoid, an N-acetylglucosamine-terminated glycoprotein, was selectively taken up in vitro by endothelial cells. Uptake was shown to be protein dependent, calcium ion dependent, and saturable, and could be described by Michaelis-Menten kinetics (apparent Km 0.29 ,M; apparent maximum velocity 4.8 pmol/h per 5 X 106 cells). Uptake was inhibited not only by Nacetylglucosamine, mannose, and mannan but also by glucose, fructose, and a glucose-albumin conjugate. Inhibition by glucose was competitive over a wide range of concentrations and was almost 100% at a glucose concentration of 56 mM. Fasting and the induction of diabetes mellitus prior to isolation of cells was associated with 60% reductions in the recovery of endothelial cells. Uptake by cells isolated from fasted rats was enhanced (apparent maximum velocity 14.3 pmol/ h per 5 X 10 cells without change in the apparent K.).These observations suggest that fasting is associated with a marked increase in the mean number of glycoprotein receptors per endothelial cell isolated from normal rats. This effect of fasting could be due to upregulation of glycoprotein receptors on endothelial cells or to the selective isolation of a subpopulation of endothelial cells from fasted animals that bears more glycoprotein receptors per cell than does another sub-

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The viability of cells grown or centrifuged in a new density gradient medium, Percoll(TM)

Cited by 215 publications

References 9 publications

Evaluation of Intracellular Killing of Bacteria by Enriched Populations of Mouse Peritoneal Exudate Neutrophils

Evaluation of Intracellular Killing of Bacteria by Enriched Populations of Mouse Peritoneal Exudate Neutrophils

Detection of Interleukin-1 and Interleukin-6 on Cryopreserved Bovine Mammary Epithelial Cells in Vitro.

Modulation of a Glycoprotein Recognition System on Rat Hepatic Endothelial Cells by Glucose and Diabetes Mellitus

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