Description
- Abstract:
- Cell sorting is a powerful tool in basic research and therapeutic enrichment. However, common cell sorting methods, such as fluorescence-activated cell sorting (FACS) and magnetic-activated cell sorting (MACS) have significant limitations, such as generally low cell yields or restriction to binary separation, respectively. To address these limitations, we developed a two-step cell sorting method called mass-added density centrifugation (MADC) to enable non-binary separation of large cell numbers based on surface protein levels. In the first MADC step (mass-adding), antibody-directed massive microparticles bind target surface proteins to modulate single-cell density proportionally to target protein level. Secondly, microparticle-laden cells are subjected to discontinuous density gradient centrifugation, whereby they separate into discrete density bands which can be isolated for downstream use. Here we demonstrate MADC’s utility for both live and fixed cell sorts of multiple cell types based on abundance of an example target protein, CD44. CD44 quantity in separated cell groups was assayed with western blots and correlated with modulated cell density. This novel sorting method enables rapid, non-binary isolation of large quantities of cells based on surface protein levels and should prove useful in both basic science and therapeutic applications.
- Notes:
- Thesis (Sc. M.)--Brown University, 2021
Citation
Sarnik, Sylvia Amalia,
"Mass-Added Density Modulation For Sorting Cells Based On Differential Surface Protein Levels"
(2021).
Biomedical Engineering Theses and Dissertations.
Brown Digital Repository. Brown University Library.
https://repository.library.brown.edu/studio/item/bdr:tz8zbejz/
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Biomedical Engineering Theses and Dissertations
Theses and Dissertations for the Biomedical Engineering department....