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KMID : 0613820090190060728
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Journal of Life Science
2009 Volume.19 No. 6 p.728 ~ p.734
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Isolation of Monocytes with High Purity and Yield from Peripheral Blood Mononuclear Cells by Flotation Density Gradient Centrifugation
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Bae Jae-Ho
Kim Sun-Hee
Kim Dong-Wan
Son Cheol-Hun
Park You-Soo
Kang Chi-Dug
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Abstract
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In this work, a simple, inexpensive and reproducible technique of flotation density gradient centrifugation was developed to isolate monocytes with high purity and yield from peripheral blood mononuclear cells (PBMC) using Histopaque solution, density and osmolarity of which were modified to 1.072 g/㎖ and 335 mOsm with phosphate-buffered saline (PBS) and sodium chloride (NaCl) solution, respectively. The average purity of monocytes was 74.75±3.84%, with the individual purity ranging from 71.44% to 82.38%. The average yield of monocytes was 32.62±11.16%, with the individual yields ranging from 21.02 to 53.63%. The monocytes isolated by floatation density gradient centrifugation could be successfully cultured into morphologically, phenotypically and functionally dendritic cells in vitro. In conclusion, the entire procedure seemed to be faster and more convenient, simple and cost-effective than other monocyte isolation methods, including plastic adherence and density gradient methods, and has the potential to be developed as a closed system for clinical scale generation of dendritic cells.
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KEYWORD
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Flotation density gradient centrifugation, monocyte isolation
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