Simultaneous preparation of mononuclear and polymorphonuclear leucocytes from horse blood on Ficoll-Hypaque medium.
Abstract: Results presented show that highly purified populations of mononuclear (MN) and polymorphonuclear (PMN) leucocytes can be obtained from horse blood by a procedure similar to that previously described for the separation of these leucocytes from human blood. This involved centrifugation of horse blood on a Ficoll-Hypaque medium with a density of 1.095 g/ml. The procedure required approximately 1 h for completion and resulted in the simultaneous preparation of MN (greater than 98% purity) and PMN (greater than 96% purity) leucocytes. Cell viability exceeded 95% and cells retained immunological functions.
Publication Date: 1980-01-01 PubMed ID: 7391588DOI: 10.1016/0022-1759(80)90100-3Google Scholar: Lookup
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- Journal Article
Summary
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The research article discusses a method to separate different types of white blood cells, specifically mononuclear and polymorphonuclear leucocytes, from horse blood using a Ficoll-Hypaque medium. This study further confirms the procedure’s efficacy, based on its high purity outputs and maintenance of cell viability and functions.
Method and Procedure
- The separation of white blood cells was done through a procedure similar to one previously used for human blood. In this process, horse blood was centrifuged using a Ficoll-Hypaque medium. Ficoll-Hypaque is a type of liquid with a particular density ideal for such separations.
- The density of the medium used for the centrifugation was 1.095 g/ml, optimal for distinguishing and separating mononuclear (MN) and polymorphonuclear (PMN) leucocytes due to their different densities.
- The entire procedure required around one hour to complete, which indicates its high efficiency.
Findings
- The procedure resulted in the simultaneous preparation of MN and PMN leucocytes, both with very high purity levels – more than 98% for MN leucocytes and more than 96% for PMN leucocytes. This suggests that the process is significantly precise and effective.
- Cell viability exceeded 95% after the procedure, meaning that most cells remained alive and healthy which further increases the reliability of the method.
- The cells retained their immunological functions. This is essential as it confirms that the separation method does not impact the cells’ ability to respond to immune challenges, maintaining their functional integrity.
Implications and Contributions
- This study significantly contributes to the field by determining a reliable method for extracting specific leucocyte types from horse blood. It may pave the way for further studies and applications requiring pure and viable blood cell samples.
- Furthermore, by confirming the method’s effectiveness on horse blood, it also suggests potential applicability for other animal species.
Cite This Article
APA
Ferrante A, Thong YH.
(1980).
Simultaneous preparation of mononuclear and polymorphonuclear leucocytes from horse blood on Ficoll-Hypaque medium.
J Immunol Methods, 34(4), 279-285.
https://doi.org/10.1016/0022-1759(80)90100-3 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Cell Separation / methods
- Ficoll
- Horses / immunology
- Microscopy
- Monocytes / classification
- Monocytes / immunology
- Neutrophils / classification
- Neutrophils / immunology
Citations
This article has been cited 4 times.- Anbukumar DS, Shornick LP, Albert CJ, Steward MM, Zoeller RA, Neumann WL, Ford DA. Chlorinated lipid species in activated human neutrophils: lipid metabolites of 2-chlorohexadecanal.. J Lipid Res 2010 May;51(5):1085-92.
- Locke LW, Chordia MD, Zhang Y, Kundu B, Kennedy D, Landseadel J, Xiao L, Fairchild KD, Berr SS, Linden J, Pan D. A novel neutrophil-specific PET imaging agent: cFLFLFK-PEG-64Cu.. J Nucl Med 2009 May;50(5):790-7.
- Ferrante A, James DW, Betts WH, Cleland LG. Rapid single-step method for purification of polymorphonuclear leucocytes from blood of patients with rheumatoid arthritis.. Clin Exp Immunol 1982 Mar;47(3):749-52.
- Zhu D, Cheng CF, Pauli BU. Blocking of lung endothelial cell adhesion molecule-1 (Lu-ECAM-1) inhibits murine melanoma lung metastasis.. J Clin Invest 1992 Jun;89(6):1718-24.
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