Optimized density gradient separation of leukocyte fractions from whole blood by adjustment of osmolarity
(1986) In Journal of Immunological Methods 93(2). p.183-191- Abstract
- Some of the compounds used for density gradient separation of blood cells have high osmolarities at the concentrations needed to create the required specific densities. Several mixed media use a combination of hyperosmolar shrinkage and red cell aggregation to improve cell separation. Due to the characteristics of Percoll density gradient medium the density and osmolarity of the gradient can be controlled separately. In the present study, Percoll gradients were used to determine the buoyant densities of different human blood cells at the osmolarities 300 mosM, 350 mosM and 400 mosM. Cell volumes were measured at the same osmolarities using a Coulter counter with channelyzer. As expected, the cell buoyant densities increased and the cell... (More)
- Some of the compounds used for density gradient separation of blood cells have high osmolarities at the concentrations needed to create the required specific densities. Several mixed media use a combination of hyperosmolar shrinkage and red cell aggregation to improve cell separation. Due to the characteristics of Percoll density gradient medium the density and osmolarity of the gradient can be controlled separately. In the present study, Percoll gradients were used to determine the buoyant densities of different human blood cells at the osmolarities 300 mosM, 350 mosM and 400 mosM. Cell volumes were measured at the same osmolarities using a Coulter counter with channelyzer. As expected, the cell buoyant densities increased and the cell volumes decreased at the higher osmolarities used. There were, however, quantitative differences between the cells with respect to the effects of an increased osmolarity, making a 350 mosM density gradient the most effective in separating mononuclear leukocytes from polymorphonuclear leukocytes. A 400 mosM gradient offered the best possibilities to separate red blood cells from polymorphonuclear leukocytes. A one-step centrifugation procedure, based on these principles, is presented. This procedure makes possible the simultaneous purification of mononuclear leukocytes and polymorphonuclear leukocytes, suitable for functional assays. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1103501
- author
- Braide, M and Bjursten, Lars Magnus LU
- organization
- publishing date
- 1986
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Density gradient, PVP silica, Buoyant density, Leukocyte separation, Osmolarity
- in
- Journal of Immunological Methods
- volume
- 93
- issue
- 2
- pages
- 183 - 191
- publisher
- Elsevier
- external identifiers
-
- pmid:3772111
- scopus:0022968624
- ISSN
- 1872-7905
- DOI
- 10.1016/0022-1759(86)90187-0
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Bioimplant Research (013242910)
- id
- d92f652b-083e-4365-b3cf-ae9eb315a2aa (old id 1103501)
- date added to LUP
- 2016-04-01 15:22:16
- date last changed
- 2021-01-03 05:52:57
@article{d92f652b-083e-4365-b3cf-ae9eb315a2aa, abstract = {{Some of the compounds used for density gradient separation of blood cells have high osmolarities at the concentrations needed to create the required specific densities. Several mixed media use a combination of hyperosmolar shrinkage and red cell aggregation to improve cell separation. Due to the characteristics of Percoll density gradient medium the density and osmolarity of the gradient can be controlled separately. In the present study, Percoll gradients were used to determine the buoyant densities of different human blood cells at the osmolarities 300 mosM, 350 mosM and 400 mosM. Cell volumes were measured at the same osmolarities using a Coulter counter with channelyzer. As expected, the cell buoyant densities increased and the cell volumes decreased at the higher osmolarities used. There were, however, quantitative differences between the cells with respect to the effects of an increased osmolarity, making a 350 mosM density gradient the most effective in separating mononuclear leukocytes from polymorphonuclear leukocytes. A 400 mosM gradient offered the best possibilities to separate red blood cells from polymorphonuclear leukocytes. A one-step centrifugation procedure, based on these principles, is presented. This procedure makes possible the simultaneous purification of mononuclear leukocytes and polymorphonuclear leukocytes, suitable for functional assays.}}, author = {{Braide, M and Bjursten, Lars Magnus}}, issn = {{1872-7905}}, keywords = {{Density gradient; PVP silica; Buoyant density; Leukocyte separation; Osmolarity}}, language = {{eng}}, number = {{2}}, pages = {{183--191}}, publisher = {{Elsevier}}, series = {{Journal of Immunological Methods}}, title = {{Optimized density gradient separation of leukocyte fractions from whole blood by adjustment of osmolarity}}, url = {{http://dx.doi.org/10.1016/0022-1759(86)90187-0}}, doi = {{10.1016/0022-1759(86)90187-0}}, volume = {{93}}, year = {{1986}}, }