Factors influencing the abundance of the side population in a human myeloma cell line.

Sui-Lin Mo, Jia Li, Yen S. Loh, Ross D. Brown, Adrian L. Smith, Yuling Chen, Douglas Joshua, Basil D. Roufogalis, George Q. Li, Kei Fan, Michelle C. H. Ng, Daniel Man-yuen Sze

Research output: Contribution to journalArticlepeer-review

Abstract

Side population (SP) refers to a group of cells, which is capable to efflux Hoechst 33342, a DNA-binding dye. SP cells exist both in normal and tumor tissues. Although SP abundance has been used as an indicator for disease prognostic and drug screening in many research projects, few studies have systematically examined the factors influencing SP analysis. In this study we aim to develop a more thorough understanding of the multiple factors involved in SP analysis including Hoechst 33342 staining and cell culture. RPMI-8226, a high SP percentage (SP%) human myeloma cell line was employed here. The results showed that SP% was subject to staining conditions including: viable cell proportion, dye concentration, staining cell density, incubation duration, staining volume, and mix interval. In addition, SP% was highest in day one after passage, while dropped steadily over time. This study shows that both staining conditions and culture duration can significantly affect SP%. In this case, any conclusions based on SP% should be interpreted cautiously. The relation between culture duration and SP% suggests that the incidence of SP cells may be related to cell proliferation and cell cycle phase. Maintaining these technical variables consistently is essential in SP research.
Original languageEnglish
Article number524845
Pages (from-to)1-8
Number of pages8
JournalBone Marrow Research
Volume2011
DOIs
Publication statusPublished - 22 Sep 2011
Externally publishedYes

Bibliographical note

Copyright the Author(s) 2011. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

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