Genome-edited human pluripotent stem cell–derived macrophages as a model of reverse cholesterol transport—brief report
Abstract
Objective—: To create isogenic human pluripotent stem cell–derived macrophages with and without ABCA1 expression as a model for reverse cholesterol transport. Approach and Results—: The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) genome-editing system was used to introduce frameshift mutations into the coding sequence of ATP-binding cassette, subfamily A, member 1. Individual human pluripotent stem cell clones with deleterious mutations were identified, expanded, and differentiated into mature macrophages with a cytokine-based, feeder-free differentiation protocol. Wild-type cells demonstrated effective cholesterol efflux to apoAI acceptor, whereas ABCA1 −/− cells displayed significantly reduced efflux ability and increased expression of proinflammatory cytokines. Conclusions—: Human pluripotent stem cell–derived macrophages capable of reverse cholesterol transport can be rapidly generated and genetically edited with CRISPR/Cas9. Introduction of homozygous frameshift mutations results in loss of ABCA1 expression in differentiated macrophages and subsequent reduction of cholesterol efflux capability. This facile genome-editing approach and differentiation protocol pave the way for future studies of the molecular determinants of reverse cholesterol transport and other macrophage properties.
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The protocol is now widely used by others.
“In 2008, we derived human PSC macrophages using a very simple methodology without OP9 feeders (Karlsson et al., 2008), which we subsequently adapted to a fully defined, robust serum-free protocol (van Wilgenburg et al., 2013), now widely used by others (Aflaki et al., 2014, Alasoo et al., 2015, Gupta et al., 2016).”
From A highly efficient human pluripotent stem cell microglia model displays a neuronal-co-culture-specific expression profile and inflammatory response (2017) · cites this paper as Gupta et al., 2016
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Induced pluripotent stem cell model recapitulates pathologic hallmarks of Gaucher disease
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Haenseler, W., Sansom, S. N., Buchrieser, J., Newey, S. E., Moore, C. S., Nicholls, F. J., Chintawar, S., Schnell, C., Antel, J. P., Allen, N. D., et al. (2017) · Stem Cell Reports · doi:10.1016/j.stemcr.2017.05.017 · Cited by 593 · see citing works
- Authors
- Gupta, R. M., Meissner, T. B., Cowan, C. A., & Musunuru, K.
- Venue
- Arteriosclerosis, Thrombosis, and Vascular Biology
- Published
- 2016-01
- Volume
- 36
- Issue
- 1
- Pages
- 15-18
- ISSN
- 1079-5642
- 1524-4636
- PMC
- PMC4690765
- Cited by
- 36
- References
- 4
- Language
- en
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