Isolation of horse mononuclear cells, especially of monocytes, on Isopaque-Ficoll neutral density gradient.
Abstract: Horse mononuclear cells were separated from whole blood using neutral density gradient centrifugation on Isopaque-Ficoll. The resulting cell suspension was comparable in composition with similarly prepared human and bovine mononuclear cell preparations. The relative concentration of monocytes was increased by the use of a gradient with density lower than that originally proposed by Böyum (Böyum, A. 1968. Scand. J. Clin. Lab. Investig. 21 supple. 97:77-89). Contamination by neutrophils was limited either by using a gradient medium of lower density or by replacing Isopaque-Ficoll by Percoll-0.9% NaCl. Although the density of the Isopaque-Ficoll appears to be the main determinant in the isolation method of Böyum, the mechanism of separation of the cell population is complex and a substantial variability of the results can be expected.
Publication Date: 1983-05-01 PubMed ID: 6306903DOI: 10.1016/0165-2427(83)90009-0Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The research involves isolating horse mononuclear cells, particularly monocytes, by using a neutral density gradient centrifugation on Isopaque-Ficoll. The method resulted in a cell suspension comparable to those derived from similar processes conducted on human and bovine cells. Experiment modifications, like using a lower density gradient and exchanging Isopaque-Ficoll with Percoll-0.9% NaCl, helped augment the concentration of monocytes and limit the neutrophil contamination respectively.
Main objectives of the research
- The main focus of the research was to isolate horse mononuclear cells, concentrating specifically on monocytes by using a process known as neutral density gradient centrifugation on Isopaque-Ficoll.
- The study also aimed to compare the resulting mononuclear cell composition with similar preparations derived from human and bovine cells.
Solution approach
- The researchers used a neutral density gradient centrifugation on Isopaque-Ficoll to separate the horse mononuclear cells from the whole blood.
- To increase the relative concentration of monocytes, the team used a gradient with lower density than the one originally proposed by Böyum.
- In order to limit neutrophil contamination, the researchers employed a gradient medium of lower density or replaced Isopaque-Ficoll with Percoll-0.9% NaCl.
Conclusion of the research
- The research concluded by stating that the resulting cell suspension from the experiment was comparable to those derived from similar processes conducted on human and bovine cells.
- They found that the density of the Isopaque-Ficoll was the main determinant in the Böyum isolation method. However, they acknowledged that the cell population’s separation mechanism was complex and that substantial variability in the results should be expected.
Cite This Article
APA
Bruyninckx WJ, Blancquaert AM.
(1983).
Isolation of horse mononuclear cells, especially of monocytes, on Isopaque-Ficoll neutral density gradient.
Vet Immunol Immunopathol, 4(4), 493-504.
https://doi.org/10.1016/0165-2427(83)90009-0 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Cell Separation / methods
- Centrifugation, Density Gradient
- Ficoll
- Horses / blood
- Lymphocytes
- Metrizoic Acid
- Monocytes
- Neutrophils
- Povidone
- Silicon Dioxide
- Sodium Chloride
Citations
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- Cossarizza A, Chang HD, Radbruch A, Akdis M, Andrä I, Annunziato F, Bacher P, Barnaba V, Battistini L, Bauer WM, Baumgart S, Becher B, Beisker W, Berek C, Blanco A, Borsellino G, Boulais PE, Brinkman RR, Büscher M, Busch DH, Bushnell TP, Cao X, Cavani A, Chattopadhyay PK, Cheng Q, Chow S, Clerici M, Cooke A, Cosma A, Cosmi L, Cumano A, Dang VD, Davies D, De Biasi S, Del Zotto G, Della Bella S, Dellabona P, Deniz G, Dessing M, Diefenbach A, Di Santo J, Dieli F, Dolf A, Donnenberg VS, Dörner T, Ehrhardt GRA, Endl E, Engel P, Engelhardt B, Esser C, Everts B, Dreher A, Falk CS, Fehniger TA, Filby A, Fillatreau S, Follo M, Förster I, Foster J, Foulds GA, Frenette PS, Galbraith D, Garbi N, García-Godoy MD, Geginat J, Ghoreschi K, Gibellini L, Goettlinger C, Goodyear CS, Gori A, Grogan J, Gross M, Grützkau A, Grummitt D, Hahn J, Hammer Q, Hauser AE, Haviland DL, Hedley D, Herrera G, Herrmann M, Hiepe F, Holland T, Hombrink P, Houston JP, Hoyer BF, Huang B, Hunter CA, Iannone A, Jäck HM, Jávega B, Jonjic S, Juelke K, Jung S, Kaiser T, Kalina T, Keller B, Khan S, Kienhöfer D, Kroneis T, Kunkel D, Kurts C, Kvistborg P, Lannigan J, Lantz O, Larbi A, LeibundGut-Landmann S, Leipold MD, Levings MK, Litwin V, Liu Y, Lohoff M, Lombardi G, Lopez L, Lovett-Racke A, Lubberts E, Ludewig B, Lugli E, Maecker HT, Martrus G, Matarese G, Maueröder C, McGrath M, McInnes I, Mei HE, Melchers F, Melzer S, Mielenz D, Mills K, Mirrer D, Mjösberg J, Moore J, Moran B, Moretta A, Moretta L, Mosmann TR, Müller S, Müller W, Münz C, Multhoff G, Munoz LE, Murphy KM, Nakayama T, Nasi M, Neudörfl C, Nolan J, Nourshargh S, O'Connor JE, Ouyang W, Oxenius A, Palankar R, Panse I, Peterson P, Peth C, Petriz J, Philips D, Pickl W, Piconese S, Pinti M, Pockley AG, Podolska MJ, Pucillo C, Quataert SA, Radstake TRDJ, Rajwa B, Rebhahn JA, Recktenwald D, Remmerswaal EBM, Rezvani K, Rico LG, Robinson JP, Romagnani C, Rubartelli A, Ruckert B, Ruland J, Sakaguchi S, Sala-de-Oyanguren F, Samstag Y, Sanderson S, Sawitzki B, Scheffold A, Schiemann M, Schildberg F, Schimisky E, Schmid SA, Schmitt S, Schober K, Schüler T, Schulz AR, Schumacher T, Scotta C, Shankey TV, Shemer A, Simon AK, Spidlen J, Stall AM, Stark R, Stehle C, Stein M, Steinmetz T, Stockinger H, Takahama Y, Tarnok A, Tian Z, Toldi G, Tornack J, Traggiai E, Trotter J, Ulrich H, van der Braber M, van Lier RAW, Veldhoen M, Vento-Asturias S, Vieira P, Voehringer D, Volk HD, von Volkmann K, Waisman A, Walker R, Ward MD, Warnatz K, Warth S, Watson JV, Watzl C, Wegener L, Wiedemann A, Wienands J, Willimsky G, Wing J, Wurst P, Yu L, Yue A, Zhang Q, Zhao Y, Ziegler S, Zimmermann J. Guidelines for the use of flow cytometry and cell sorting in immunological studies. Eur J Immunol 2017 Oct;47(10):1584-1797.
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