Direkt zum Inhalt
Merck
HomeMammalian Cell CultureApplications of Percoll In Other Cell Types

Applications of Percoll in Other Cell Types

The following tables were compiled to assist the researcher in selecting references most likely to contain relevant information regarding use of Percoll for a particular cell or tissue type.

Liver cells

SpeciesGradient typeTissue typeCommentsDownstream applicationRef. #
humancontinuousliverPurification of cryo-preserved hepatocytes on Percoll density gradients increased the percentage of viable cells from 55 to 87%.primary cell culture, electron microscopy, viability assay radiolabeled protein synthesis, secretion assay, metabolic studies, toxicological studies975
ratcontinuousliverPercoll offered a good way to obtain an enriched population of Kupffer cells. Recovery was 82%, viability 87% and purity 67%.peroxidatic reaction20
ratcontinuousliverPercoll gradients were used to isolate hepatocyte plasma membranes and mitochondrial membranes.phase contrast microscopy, cell binding experiments33
ratcontinuousliverRat liver cells furnished subpopulations of parenchymal cells (hepatocytes) having buoyant densities of 1.07 to 1.09 g/mL, and non-parenchymal cells (mostly phagocytosing Kupffer cells) at a density of 1.04 to 1.06 g/mL.cell culture55
ratNAliverFinal preparations contained less than 5% nonviable cells as judged by trypan blue exclusion.cell culture71
ratcontinuousliverPercoll gradients were used to franctionate nonparenchymal cells into Kupffer cells, stellate and endothelial cells.light and flourescence microscopy, carboxyesterase and Glutathione- S-transferase (GST) activities976
ratdiscontinuous (2-layer)liverPercoll provided a simple, low cost, and rapid method for the isolation, purification and cultivation of rat liver sinusoidal endothelial cells (LEC).electron microscopy, cell culture, trypan blue exclusion977
ratdiscontinuous (2-step)liverPercoll gradients were used to separate fat storing cells (FSC) from liver endothelial cells (LEC) and Kupffer cells (KC).cell culture978
ratcontinuousliverFollowing the removal of damaged cells by centrifugation in Percoll, the mean viability of cryo-preserved hepatocytes, tested by trypan blue exclusion, was 88.6% (±1.3%).cell viability and study of xenobiotic metabolism979
ratcontinuousliverPercoll was used to remove dead cells from cryopreserved cells. Cell viability was 88 ±1% after the Percoll step.cell viability and study of xenobiotic metabolism980
ratcontinuousliverIf cryo-preserved cells were purified by a Percoll centrifugation after thawing, the enzyme activities were not significantly different from those of freshly isolated parenchymal cells, and the viability was 86%.Lowry protein assay, cytochrome assay, enzyme assays981
ratcontinuousliverPercoll separation yielded cryo-preserved cells with a viability and metabolic capacity not measurably different from freshly isolated cells.protein determination, enzyme assays and metabolism of testosterone and benzo(a) pyrene (BaP)982
ratdiscontinuous (2-layer)liverPercoll two-step gradients were used to separate Kupffer cells (KC) and liver endothelial cells (LEC). Preparations of KC were 85 to 92% homogenous while the LEC preparation was at least 95% pure.light microscopy, electron microscopy and peroxidase staining983
ratdiscontinuous (5-layer)liver, spleenPercoll gradients were used to separate both spleen and liver cells. Spleen and liver cell viability was over 95%.trypan blue viability assay, cell culture984
ratcontinuousliver biopsyPercoll was used for separation of hepatocytes and non-parenchymal cells, as well as subfractionation.cell enumeration using Coulter counter, immunocytochemistry, DNA extraction, Southern blot analysis, assay of marker enzymes and protein in subcellular fractions, electron microscopy985

Leydig cells

SpeciesGradient typeTissue typeCommentsDownstream applicationRef. #
humancontinuoustestisPercoll-purified Leydig cells were 70 to 80% pure based on staining for 3 beta-hydroxysteroid dehydrogenase.cell culture, stimulation of testosterone production986
humancontinuoustestisPercoll-purified Leydig cells were 80 to 90% pure as determined by 3 betahydroxysteroid Dehydrogenase staining.immunocytochemical localization of apolipoprotein E (apoE)987
humandiscontinuous (4-layer)testisPercoll gradients were used to isolate human Leydig cell mesenchymal precursors.cell culture988
humandiscontinuous (5-layer)testisPercoll gradient centrifugation permitted isolation of two Leydig cell fractions.cell culture989
mousecontinuous (linear)testisTwo groups were obtained: group 1 had densities of 1.0667 to 1.0515 g/mL; group 2 had densities of 1.0514 to 1.0366 g/mL.in vitro testosterone production electron microscope stereology990
porcinediscontinuoustestisPurity of Leydig cells was > 85%.effect of hydrocortisone (HS) and adrenocorticotropic hormone (ACTH) on testosterone production991
ratcontinuoustestisRat Leydig cells were purified from testis using elutriation followed by Percoll gradient centrifugation. cell culture, the effect of human chorionic gonadotropin (hCG) on its gene regulation and protein secretion992
ratcontinuoustestis cell culture, the effect of GHreleasing hormone (GHRH) on Leydig cell steroidogensis993
ratcontinuoustestisRat Leydig cells were purified from testis using elutriation followed by Percoll gradient centrifugation. Band 2(of 3) contained > 95% Leydig cells (average density was 1.075 g/mL). cell culture in presence of 125Ilabeled hCG, testosterone and cAMP production994
ratcontinuoustestisComparison of Leydig cells of different densities were made.viability staining, cell culture995
ratcontinuoustestis viability staining, in vitro testosterone production, SDSPAGE electrophoresis996
ratcontinuoustestisIsolation by Percoll gradient resulted in complete retention of morphological and biological integrity and a purity of 90 to 95%.cell culture in presence of human chorionic gonadotropin (hCG), phase contrast microscopy, light microscopy and electron microscopy31
ratdiscontinuous (2-step)testis cell culture in the presence of interleukin-1 (IL-1)997
ratcontinuous (selfgenerating)testisLeydig cell precursors and pure (96%) Leydig cells were isolated on Percoll gradients.cell culture in presence of human chorionic gonadotropin (hCG)998
ratdiscontinuoustestisThe purity of Leydig cells ranged from 90 to 95%.cell culture in presence of human chorionic gonadotropin (hCG)999
ratdiscontinuous and continuoustestisIn the discontinuous gradient, the densest fraction contained a high proportion of Leydig cells whereas the lighter fraction contained mostly non-Leydig cells.125I-labeled iododeoxyuridine incorporation1000

Spermatozoa

SpeciesGradient typeCommentsDownstream applicationRef. #
bovinediscontinuousPercoll was thought to improve semen and preserve acrosome integrity.acrosome microscopy evaluation1024
hamstercontinuousCaput epididymal spermatoazoa, with a specific gravity of 1.10-1.12 g/mL, were isolated without contamination by other cells.lipid extraction and fractionation electron microscopy1025
macaquecontinuousPercoll separation resulted in increased spermzona binding and did not affect the percentage of acrosome-reacted sperm bound to the zona or the percent motility and percentage of acrosome-reacted sperm in suspension.zona binding experiments, acrosome reaction, motility assays1026

Bone marrow cells

SpeciesGradient typeTissue typeCommentsDownstream applicationRef. #
normal humandiscontinuous (2-layer)bone marrowMegakaryocytes were at the interface between 1.020 g/mL and 1.050 g/mL.magnetic beads for further purification, flow cytometry1027
normal humandiscontinuousbloodB cells were recovered at least 95% pure. Gradients removed B-cell blasts very effectively.flow cytometry1028
HIV infected, normal and immune throm-bocytopenic purpura humandiscontinuous (2-layer)bone marrowCells at the 1.020/1.050 interface were enriched 10-fold in megakaryocytes, while those at the 1.050/1.070 interface were immature cells. megakaryocyte cultures prepared from immature cells for in situ hybridization1029
normal humandiscontinuous (2-layer)bone marrowPercoll density fractionation resulted in the depletion of greater than 95% of total marrow cells and an increase in megakaryocyte frequency from about 0.05% to 3 to 7%.preparation of RNA and subsequent PCR, flow cytometry1030
normal and arthritic humandiscontinuous (3-layer)bone marrowCells prepared were suitable for cell culture.colony plaque assay, immunoflourscence, flow cytometry, protein colony blotting, RNA-colony blotting1031
normal and leukemic humandiscontinuous (4-layer)peripheral bloodLow density cells post- and pre-transplant were prepared for analysis.magnetic beads for further purification, PCR1032
normal humandiscontinuous (7-layer)bone marrowT cells obtained using Percoll were enriched about two-fold in the high- density fractions of marrow cells and depleted by about four- to five-fold in the lowest-density fraction as compared with Ficoll™.flow cytometry, mixed lymphocyte reaction assay, natural killer cell assay, cell culture1033
normal humandiscontinuous (1-layer)bone marrowBone marrow cells were prepared using Percoll to remove RBC. isolation of CD34+ cells using soybean agglutinin-coated flasks, progenitor cell assays, and flow cytometry1034
marmosetdiscontinuous (1-layer)bone marrowBone marrow megakaryocytes from both interleukin-6 (IL-6) treated and untreated animals could be separated in Percoll.flow cytometry1035
primatediscontinuous (1-layer)bone marrowBone Marrow was isolated from both normal monkeys and interleukin-6 (IL-6) treated monkeys.cell enumeration, FACS, digital imaging microscopy and electron microscopy1036
monkeydiscontinuous (1-layer)bone marrow peripheral and bloodLight density cells were prepared from aspirates over a 60% cushion.cell culture and identification of various colony types1037
mousediscontinuous (1-layer)bone marrowRed blood cells were removed from bone-marrow preparations with a single 70% Percoll cushion.culture of hematopoietic precursers, effects of interleukin-10 (IL-10) on proliferation, alkaline phosphatase activity, collagen synthesis assay, osteocalcin, preparation of RNA, and electron microscopy1038
mousediscontinuous (3-layer)bone marrowBone marrow progenitor cells were suitable for culture.effects of interleukin-3 (IL-3) and lipoplysaccharide (LPS) on cultured cells1039
mousediscontinuous (3-layer)bone marrowCells prepared were depleted of lymphoid and macrophage-lineage cells by addition of monoclonal antibody plus complement.FACS analysis, hematopoietic progenitor cell culture, reconstitution of lethally irradiated mice1040
mousediscontinuous (3-layer)bone marrowPercoll was used to separate bone marrow fractions containing mostly blasts and lymphoid cells from those containing a high level of colony-forming units-spleen (CFU-S) counts.FACS analysis, chemotaxis assay, assay of colony-forming units-spleen (CFU-S)1041
mousediscontinuous (3-layer)proteasetreated calvarial bone sectionsPercoll gradients gave distinct subpopulations of cells based upon the results of various assays.primary cell culture, flow cytometry, insulin-like growth factor I (IGF-I) assay, binding of epidermal growth factor, alkaline phosphatase determination1042
mousediscontinuous (4-layer)bone marrowNormal suppressor cell activity was maintained after separation. suppressor cell activity assay1043
mousediscontinuous (4-layer)bone marrowCells at a 1.06/1.07 g/mL density were used in subsequent studies.reconstitution of lethally irradiated animals1044
mousediscontinuous (5-layer)bone marrow, spleen flow cytometry, reconstitution of lethally irradiated mice1045
ratdiscontinuous (3-layer)bone marrowAbout 75% of the input CFUmegakaryocytes (CFU-MK) were recovered in the fraction between 1.063 and 1.082 g/mL Percoll. CFU-MK were enriched only in this density fraction.culture of hematopoietic progenitor cells1046
rabbitcontinuousbone marrow implantation into in vivo placed diffusion chamber, cytochemical staining, and electron microscopy 38
felinediscontinuous (1-layer)bone marrowMarrow mononuclear cells from both feline immunodeficiency virus-infected cats and normal cats were isolated.culture of hematopoietic progenitor cells1047

Macrophages

SpeciesGradient typeTissue typeCommentsDownstream applicationRef. #
humandiscontinuouslungAlveolar macrophages were purified from contaminating granulocytes using a discontinuous Percoll gradient.superoxide (SO) release1048
humandiscontinuous (4-layer)brochoalveolar lavagePercoll gradients gave > 95% alveolar macrophage (AM) purity.cell viability assay, light microscopy1049
humandiscontinuous (4-layer)lungUse of Percoll resulted in near total purification of alveolar macro-phages (AM) from other cells.superoxide (SO) anion release1050
humandiscontinuous (4-layer)decidual tissueWhen cells were purified further with Percoll, the percentage of CD-14- positive cells increased by 52%.secretion of plateletactivating factor (PAF) acetylhydrolase1051
humandiscontinuouspulmonary> 97% of the cells of fractions 1 to 4 were (4-layer) shown to be alveolar macro-phages (AM) in a previous study.nonspecific esterase staining, flow cytometric DNA analysis1052
humandiscontinuous (4-layer)lungThis method was used to study alveolar macrophage (AM) heterogeneity. The increased numbers of hypodense AM found in the asthmatic patients were unlikely to be due to the procedure.  cell viability, esterase and peroxidase activity assays, electron microscopy, generation of superoxide anion and thromboxane B21053
humandiscontinuous (5-layer)peripheral bloodPercoll-isolated monocyte/macrophages were harvested from the top layer and routinely contained 75/90% monocytes/macrophages as identified by Wright-Giemsa stain.interactions between monocyte/macrophage and vascular smooth muscle cells928
mousecontinuous and discontinuousperitoneumThe total cell yield was 100.0% ±0.8%, and as measured by the trypan blue exlusion test, the cell viability was completely preserved.  light microscopy, trypan blue exclusion, esterase activity assay, peroxidase activity assay, cell immunophenotyping, bacterial phagocytic assays1054
mousediscontinuous (4-layer)cultured cellsPercoll did not have a detectable effect on the cytolytic activity of cultured macrophages or on their viability.phagocytic and cytolytic assays30
mouse, ratcontinuous and discontinuousperitoneumA continuous gradient followed by a discontinuous gradient was used to isolate all cell populations according to their actual density. This procedure yielded cells of high viability with preservation of critical cell functiontrypan blue exclusion1055
ratdiscontinuous (5-layer)lungThe Percoll fractions were designated I to IV in order of increasing density with a percent distribution of cells of about 5, 15, 50 and 30%, respectively. Cell viability was > 95%.  fluorescence microscopy1056
ratdiscontinuous (5-layer)lungCell viability was > 95% by trypan blue exclusion and > 95% were identified as alveolar macrophages (AM) in un-fractionated and fractionated cells by Giemsa and nonspecific esterase stains.effects of pulmonary surfactant and protein A on phagocytosis, light microscopy1057
ratcontinuousbronchoalveolar lavageThe various fractions comprised approximately 90 to 99% macrophages in virtually all instances.esterase activity, surface expansion of Ia antigen by an immunoperoxidase technique1058

Mast cells

SpeciesGradient typeTissue typeCommentsDownstream applicationRef. #
mouseNAperitoneumPurity of the mast cells was nearly 100%, as checked by Memacolor fast staining.qualitative and quantitative PCR analysis1059
mousecontinuousperitoneumStarting from a peritoneal cell population containing 4% mast cells, a mast cell purification of up to 95% was obtained.electron microscopy and ultrastructural cytochemistry8
ratdiscontinuousperitoneumMast cell purity with Percoll was > 95%.direct interaction between mast and non-mast cells, histamine release assay1060
ratcontinuousperitoneumMast cells purified on Percoll gradients were more than 90% pure by toluidine blue staining, and the viability was > 98% by the trypan blue exclusion test.flourometric assay to measure histamine release1061
ratcontinuousperitoneumMast cells can be isolated with high yields and purity by centrifugation on gradients of Percoll.light and electron microscopy, cytofluorometry9
ratcontinuous (sequential)peritoneumThe purity of mast cells purified over sequential Percoll gradients was evaluated by measurement of the contribution of eosinophil peroxidase to mast cell peroxidase activity.histamine release and peroxidase activity1062

Thymocytes

SpeciesGradient typeTissue typeCommentsDownstream applicationRef. #
mousediscontinuous (5-layer)thymusPercoll was used for separation of immature thymocytes.in vitro stimulation by mitogens, isolation of nuclei, isolation and gel electrophoresis DNA, enzyme assays1063
ratdiscontinuousthymusPercoll was used for separation of normal and apoptotic thymocytes.flow cytometry1064
ratdiscontinuous (4-layer)thymusPercoll was used for separation of cells possessing the characteristically condensed nuclear chromatin associated with apoptosis from apparently normal thymocytes.electron microscopy, Coulter counter analysis, flow cytometry, DNA analysis1065
ratdiscontinuous (4-layer)thymusPercoll was used for isolation of a transitional population of preapoptotic thymocytes.DNA analysis, isolation of nuclei and DNA autodigestion, light and electron microscopy1066
rat, mousediscontinuous (3-layer)thymusPercoll was used to separate large and small thymocytes. An extremely high level of viability was maintainedphase contrast microscopy and autoradiography62

Miscellaneous cells

Cell TypeSpeciesGradient typeTissue typeCommentsDownstream applicationRef. #
pancreatic isletshuman, mousecontinuouspancreasThe use of Percoll eliminated the problems of high viscosity, undesired osmotic properties and, in some cases, also toxic effects.density determination and insulin secretion5
endothelialhumancontinuous linear gradientwhole bloodFinal recovery of endothelial cells was 91.6%.immunofluorescence1067
trophoblastsratcontinuousplacentaPercoll gradient centrifugation yielded efficient separation of rat placental lactogen- II (rPL-II) producing cells from digested tissue from labyrinth and junctional zones of the chorioallantoic placenta.development of in vitro rat placental trophoblast cell culture system1068
variousNANANAThis paper compared different approaches to cell separation. According to the authors, Percoll is generally the most useful media for isopycnic centrifugation of most kinds of cells.none1069
viable vs. nonviablehuman, ratdiscontinuous (2-layer)various tumor tissueInterface showed a viability of > 90%, but the yield of viable cells decreased dramatically if the tissue resection was not immediately processed.trypan blue viability assay, 2-D PAGE1070
apoptotichumandiscontinuous (7-layer)promyelocytic leukemic cell lineThe step gradient used generated three main cell bands and a cell pellet, the pellet was very enriched for apoptotic cells (85 to 90%).DNA isolation1071
lymphoblasthumancontinuouswhole bloodLymphoblasts were enucleated using a Percoll gradient containing cytochalasin B.electrofusion1072
brain capillary endothelialratcontinuous pre-madebrainSubsequent Percoll gradient centrifugation resulted in a homogenous population of capillary endothelial cells capable of attachment to collagen and incorporation of tritiated thymidine.cell culture, light microscopy electron microscopy170
neuronsrabbitdiscontinuous and rate zonaldorsal-root gangliaNeurons were isolated with a viability of 80% and a purity of > 90%.cell culture, light and electron microscopy480
nonmyogenic separated from myogenicchickendiscontinuousbreast musclesSeparation of cells from embryonic muscle allowed direct analysis of cell-specific proteins without the need for cell culturing.cell culture, microscopy, DNA/ protein analysis680
megakaryocyteshumandiscontinuousbone marrowIsolation of megakaryocytes was reproducibly better in Percoll than in BSA.Ficoll 400 centrifugation to further purify, complement receptor assay155
chondrocytesratdiscontinuousbone marrowCell viability was > 95% while yield varied depending on aggregation of cells.cell culture, quantitation of proteoglycans and collagen629
spermiophagesturkeydiscontinuousspermSpermiophages fixed immediately after Percoll isolation resembled those in freshly ejaculated semen except for an apparent increase in the number of mitochondria.light and electron microscopy, cell culture1073
NAhumancontinuousparathyroid glandDensities of parathyroid glands were measured using various density gradient media. For densities > 1.0 g/mL, Percoll proved superior to any of the other gradient liquids investigated.glandular density determination2
Materials
Produkt-Nr.ProduktbezeichnungBeschreibungPreisangaben
GE17-0300-05Ficoll® PM400Cytiva 17-0300-05, pack of 5 kg
Melden Sie sich an, um fortzufahren.

Um weiterzulesen, melden Sie sich bitte an oder erstellen ein Konto.

Sie haben kein Konto?