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 | |||||
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Species | Gradient type | Tissue type | Comments | Downstream application | Ref. # |
human | continuous | liver | Purification 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 studies | 975 |
rat | continuous | liver | Percoll offered a good way to obtain an enriched population of Kupffer cells. Recovery was 82%, viability 87% and purity 67%. | peroxidatic reaction | 20 |
rat | continuous | liver | Percoll gradients were used to isolate hepatocyte plasma membranes and mitochondrial membranes. | phase contrast microscopy, cell binding experiments | 33 |
rat | continuous | liver | Rat 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 culture | 55 |
rat | NA | liver | Final preparations contained less than 5% nonviable cells as judged by trypan blue exclusion. | cell culture | 71 |
rat | continuous | liver | Percoll gradients were used to franctionate nonparenchymal cells into Kupffer cells, stellate and endothelial cells. | light and flourescence microscopy, carboxyesterase and Glutathione- S-transferase (GST) activities | 976 |
rat | discontinuous (2-layer) | liver | Percoll 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 exclusion | 977 |
rat | discontinuous (2-step) | liver | Percoll gradients were used to separate fat storing cells (FSC) from liver endothelial cells (LEC) and Kupffer cells (KC). | cell culture | 978 |
rat | continuous | liver | Following 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 metabolism | 979 |
rat | continuous | liver | Percoll was used to remove dead cells from cryopreserved cells. Cell viability was 88 ±1% after the Percoll step. | cell viability and study of xenobiotic metabolism | 980 |
rat | continuous | liver | If 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 assays | 981 |
rat | continuous | liver | Percoll 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 |
rat | discontinuous (2-layer) | liver | Percoll 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 staining | 983 |
rat | discontinuous (5-layer) | liver, spleen | Percoll gradients were used to separate both spleen and liver cells. Spleen and liver cell viability was over 95%. | trypan blue viability assay, cell culture | 984 |
rat | continuous | liver biopsy | Percoll 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 microscopy | 985 |
Leydig cells | |||||
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Species | Gradient type | Tissue type | Comments | Downstream application | Ref. # |
human | continuous | testis | Percoll-purified Leydig cells were 70 to 80% pure based on staining for 3 beta-hydroxysteroid dehydrogenase. | cell culture, stimulation of testosterone production | 986 |
human | continuous | testis | Percoll-purified Leydig cells were 80 to 90% pure as determined by 3 betahydroxysteroid Dehydrogenase staining. | immunocytochemical localization of apolipoprotein E (apoE) | 987 |
human | discontinuous (4-layer) | testis | Percoll gradients were used to isolate human Leydig cell mesenchymal precursors. | cell culture | 988 |
human | discontinuous (5-layer) | testis | Percoll gradient centrifugation permitted isolation of two Leydig cell fractions. | cell culture | 989 |
mouse | continuous (linear) | testis | Two 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 stereology | 990 |
porcine | discontinuous | testis | Purity of Leydig cells was > 85%. | effect of hydrocortisone (HS) and adrenocorticotropic hormone (ACTH) on testosterone production | 991 |
rat | continuous | testis | Rat 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 secretion | 992 |
rat | continuous | testis | cell culture, the effect of GHreleasing hormone (GHRH) on Leydig cell steroidogensis | 993 | |
rat | continuous | testis | Rat 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 production | 994 |
rat | continuous | testis | Comparison of Leydig cells of different densities were made. | viability staining, cell culture | 995 |
rat | continuous | testis | viability staining, in vitro testosterone production, SDSPAGE electrophoresis | 996 | |
rat | continuous | testis | Isolation 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 microscopy | 31 |
rat | discontinuous (2-step) | testis | cell culture in the presence of interleukin-1 (IL-1) | 997 | |
rat | continuous (selfgenerating) | testis | Leydig cell precursors and pure (96%) Leydig cells were isolated on Percoll gradients. | cell culture in presence of human chorionic gonadotropin (hCG) | 998 |
rat | discontinuous | testis | The purity of Leydig cells ranged from 90 to 95%. | cell culture in presence of human chorionic gonadotropin (hCG) | 999 |
rat | discontinuous and continuous | testis | In 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 incorporation | 1000 |
Spermatozoa | ||||
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Species | Gradient type | Comments | Downstream application | Ref. # |
bovine | discontinuous | Percoll was thought to improve semen and preserve acrosome integrity. | acrosome microscopy evaluation | 1024 |
hamster | continuous | Caput 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 microscopy | 1025 |
macaque | continuous | Percoll 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 assays | 1026 |
Bone marrow cells | |||||
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Species | Gradient type | Tissue type | Comments | Downstream application | Ref. # |
normal human | discontinuous (2-layer) | bone marrow | Megakaryocytes were at the interface between 1.020 g/mL and 1.050 g/mL. | magnetic beads for further purification, flow cytometry | 1027 |
normal human | discontinuous | blood | B cells were recovered at least 95% pure. Gradients removed B-cell blasts very effectively. | flow cytometry | 1028 |
HIV infected, normal and immune throm-bocytopenic purpura human | discontinuous (2-layer) | bone marrow | Cells 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 hybridization | 1029 |
normal human | discontinuous (2-layer) | bone marrow | Percoll 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 cytometry | 1030 |
normal and arthritic human | discontinuous (3-layer) | bone marrow | Cells prepared were suitable for cell culture. | colony plaque assay, immunoflourscence, flow cytometry, protein colony blotting, RNA-colony blotting | 1031 |
normal and leukemic human | discontinuous (4-layer) | peripheral blood | Low density cells post- and pre-transplant were prepared for analysis. | magnetic beads for further purification, PCR | 1032 |
normal human | discontinuous (7-layer) | bone marrow | T 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 culture | 1033 |
normal human | discontinuous (1-layer) | bone marrow | Bone marrow cells were prepared using Percoll to remove RBC. | isolation of CD34+ cells using soybean agglutinin-coated flasks, progenitor cell assays, and flow cytometry | 1034 |
marmoset | discontinuous (1-layer) | bone marrow | Bone marrow megakaryocytes from both interleukin-6 (IL-6) treated and untreated animals could be separated in Percoll. | flow cytometry | 1035 |
primate | discontinuous (1-layer) | bone marrow | Bone Marrow was isolated from both normal monkeys and interleukin-6 (IL-6) treated monkeys. | cell enumeration, FACS, digital imaging microscopy and electron microscopy | 1036 |
monkey | discontinuous (1-layer) | bone marrow peripheral and blood | Light density cells were prepared from aspirates over a 60% cushion. | cell culture and identification of various colony types | 1037 |
mouse | discontinuous (1-layer) | bone marrow | Red 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 microscopy | 1038 |
mouse | discontinuous (3-layer) | bone marrow | Bone marrow progenitor cells were suitable for culture. | effects of interleukin-3 (IL-3) and lipoplysaccharide (LPS) on cultured cells | 1039 |
mouse | discontinuous (3-layer) | bone marrow | Cells 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 mice | 1040 |
mouse | discontinuous (3-layer) | bone marrow | Percoll 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 |
mouse | discontinuous (3-layer) | proteasetreated calvarial bone sections | Percoll 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 determination | 1042 |
mouse | discontinuous (4-layer) | bone marrow | Normal suppressor cell activity was maintained after separation. | suppressor cell activity assay | 1043 |
mouse | discontinuous (4-layer) | bone marrow | Cells at a 1.06/1.07 g/mL density were used in subsequent studies. | reconstitution of lethally irradiated animals | 1044 |
mouse | discontinuous (5-layer) | bone marrow, spleen | flow cytometry, reconstitution of lethally irradiated mice | 1045 | |
rat | discontinuous (3-layer) | bone marrow | About 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 cells | 1046 |
rabbit | continuous | bone marrow | implantation into in vivo placed diffusion chamber, cytochemical staining, and electron microscopy | 38 | |
feline | discontinuous (1-layer) | bone marrow | Marrow mononuclear cells from both feline immunodeficiency virus-infected cats and normal cats were isolated. | culture of hematopoietic progenitor cells | 1047 |
Macrophages | |||||
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Species | Gradient type | Tissue type | Comments | Downstream application | Ref. # |
human | discontinuous | lung | Alveolar macrophages were purified from contaminating granulocytes using a discontinuous Percoll gradient. | superoxide (SO) release | 1048 |
human | discontinuous (4-layer) | brochoalveolar lavage | Percoll gradients gave > 95% alveolar macrophage (AM) purity. | cell viability assay, light microscopy | 1049 |
human | discontinuous (4-layer) | lung | Use of Percoll resulted in near total purification of alveolar macro-phages (AM) from other cells. | superoxide (SO) anion release | 1050 |
human | discontinuous (4-layer) | decidual tissue | When cells were purified further with Percoll, the percentage of CD-14- positive cells increased by 52%. | secretion of plateletactivating factor (PAF) acetylhydrolase | 1051 |
human | discontinuous | pulmonary | > 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 analysis | 1052 |
human | discontinuous (4-layer) | lung | This 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 B2 | 1053 |
human | discontinuous (5-layer) | peripheral blood | Percoll-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 cells | 928 |
mouse | continuous and discontinuous | peritoneum | The 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 assays | 1054 |
mouse | discontinuous (4-layer) | cultured cells | Percoll did not have a detectable effect on the cytolytic activity of cultured macrophages or on their viability. | phagocytic and cytolytic assays | 30 |
mouse, rat | continuous and discontinuous | peritoneum | A 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 function | trypan blue exclusion | 1055 |
rat | discontinuous (5-layer) | lung | The 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 microscopy | 1056 |
rat | discontinuous (5-layer) | lung | Cell 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 microscopy | 1057 |
rat | continuous | bronchoalveolar lavage | The various fractions comprised approximately 90 to 99% macrophages in virtually all instances. | esterase activity, surface expansion of Ia antigen by an immunoperoxidase technique | 1058 |
Mast cells | |||||
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Species | Gradient type | Tissue type | Comments | Downstream application | Ref. # |
mouse | NA | peritoneum | Purity of the mast cells was nearly 100%, as checked by Memacolor fast staining. | qualitative and quantitative PCR analysis | 1059 |
mouse | continuous | peritoneum | Starting from a peritoneal cell population containing 4% mast cells, a mast cell purification of up to 95% was obtained. | electron microscopy and ultrastructural cytochemistry | 8 |
rat | discontinuous | peritoneum | Mast cell purity with Percoll was > 95%. | direct interaction between mast and non-mast cells, histamine release assay | 1060 |
rat | continuous | peritoneum | Mast 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 release | 1061 |
rat | continuous | peritoneum | Mast cells can be isolated with high yields and purity by centrifugation on gradients of Percoll. | light and electron microscopy, cytofluorometry | 9 |
rat | continuous (sequential) | peritoneum | The 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 activity | 1062 |
Thymocytes | |||||
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Species | Gradient type | Tissue type | Comments | Downstream application | Ref. # |
mouse | discontinuous (5-layer) | thymus | Percoll was used for separation of immature thymocytes. | in vitro stimulation by mitogens, isolation of nuclei, isolation and gel electrophoresis DNA, enzyme assays | 1063 |
rat | discontinuous | thymus | Percoll was used for separation of normal and apoptotic thymocytes. | flow cytometry | 1064 |
rat | discontinuous (4-layer) | thymus | Percoll 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 analysis | 1065 |
rat | discontinuous (4-layer) | thymus | Percoll was used for isolation of a transitional population of preapoptotic thymocytes. | DNA analysis, isolation of nuclei and DNA autodigestion, light and electron microscopy | 1066 |
rat, mouse | discontinuous (3-layer) | thymus | Percoll was used to separate large and small thymocytes. An extremely high level of viability was maintained | phase contrast microscopy and autoradiography | 62 |
Miscellaneous cells | ||||||
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Cell Type | Species | Gradient type | Tissue type | Comments | Downstream application | Ref. # |
pancreatic islets | human, mouse | continuous | pancreas | The use of Percoll eliminated the problems of high viscosity, undesired osmotic properties and, in some cases, also toxic effects. | density determination and insulin secretion | 5 |
endothelial | human | continuous linear gradient | whole blood | Final recovery of endothelial cells was 91.6%. | immunofluorescence | 1067 |
trophoblasts | rat | continuous | placenta | Percoll 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 system | 1068 |
various | NA | NA | NA | This 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. | none | 1069 |
viable vs. nonviable | human, rat | discontinuous (2-layer) | various tumor tissue | Interface 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 PAGE | 1070 |
apoptotic | human | discontinuous (7-layer) | promyelocytic leukemic cell line | The step gradient used generated three main cell bands and a cell pellet, the pellet was very enriched for apoptotic cells (85 to 90%). | DNA isolation | 1071 |
lymphoblast | human | continuous | whole blood | Lymphoblasts were enucleated using a Percoll gradient containing cytochalasin B. | electrofusion | 1072 |
brain capillary endothelial | rat | continuous pre-made | brain | Subsequent 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 microscopy | 170 |
neurons | rabbit | discontinuous and rate zonal | dorsal-root ganglia | Neurons were isolated with a viability of 80% and a purity of > 90%. | cell culture, light and electron microscopy | 480 |
nonmyogenic separated from myogenic | chicken | discontinuous | breast muscles | Separation of cells from embryonic muscle allowed direct analysis of cell-specific proteins without the need for cell culturing. | cell culture, microscopy, DNA/ protein analysis | 680 |
megakaryocytes | human | discontinuous | bone marrow | Isolation of megakaryocytes was reproducibly better in Percoll than in BSA. | Ficoll 400 centrifugation to further purify, complement receptor assay | 155 |
chondrocytes | rat | discontinuous | bone marrow | Cell viability was > 95% while yield varied depending on aggregation of cells. | cell culture, quantitation of proteoglycans and collagen | 629 |
spermiophages | turkey | discontinuous | sperm | Spermiophages 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 culture | 1073 |
NA | human | continuous | parathyroid gland | Densities 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 determination | 2 |
Materials
Nº do produto | Nome do produto | Descrição | Preços |
---|---|---|---|
GE17-0300-05 | Ficoll® PM400 | Cytiva 17-0300-05, pack of 5 kg |
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