ArticleJournal of biomolecular techniques : JBT2012
Development of the "Three-step MACS": a novel strategy for isolating rare cell populations in the absence of known cell surface markers from complex animal tissue.
Article in Journal of biomolecular techniques : JBT, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 27 citations in OpenAlex.
- Liquid biopsy - a narrative review with an update on current US governmental clinical trials targeting immunotherapy.Future science OA · 2025Review
- CD52/FLAG and CD52/HA Fusion Proteins as Novel Magnetic Cell Selection Markers.International journal of molecular sciences · 2024Article
- Selection of Cell Populations with High or Low Surface Marker Expression Using Magnetic Sorting.Cells · 2023Article
- Review
- Liquid biopsy: a step closer to transform diagnosis, prognosis and future of cancer treatments.Molecular cancer · 2022Review
- A Rapid Protocol for Direct Isolation of Osteoclast Lineage Cells from Mouse Bone Marrow.Bio-protocol · 2022Article
- Advantages, Factors, Obstacles, Potential Solutions, and Recent Advances of Fish Germ Cell Transplantation for Aquaculture-A Practical Review.Animals : an open access journal from MDPI · 2022Review
- Using mass cytometry for the analysis of samples of the human airways.Frontiers in immunology · 2022Article
- Strategies for enrichment of circulating tumor cells.Translational cancer research · 2020Review
- Expression ofFrontiers in cellular neuroscience · 2018Article
- Pinocembrin protects hemorrhagic brain primarily by inhibiting toll-like receptor 4 and reducing M1 phenotype microglia.Brain, behavior, and immunity · 2017Article
- Review
- PSA-NCAM-negative neural crest cells emerging during neural induction of pluripotent stem cells cause mesodermal tumors and unwanted grafts.Stem cell reports · 2015Article
- Getting Down to Specifics: Profiling Gene Expression and Protein-DNA Interactions in a Cell Type-Specific Manner.Advances in genetics · 2015Review
- Article
- Subretinal transplantation of MACS purified photoreceptor precursor cells into the adult mouse retina.Journal of visualized experiments : JoVE · 2014Article
- Antibody-free magnetic cell sorting of genetically modified primary human CD4+ T cells by one-step streptavidin affinity purification.PloS one · 2014Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
To circumvent the difficulty of isolating specific cell populations by MACS from dissociated complex animal tissue, when their proportions reached levels similar to that of the background, we developed the "Three-step MACS" strategy. Cells of interest are defined by their expression of a particular gene(s) of interest rather by than their natural cell surface markers or size. A two-component transgenic cell surface protein, for two sequential rounds of MACS, is expressed under the promoter control of the endogenous gene of interest by means of gene targeting and the generation of transgenic tissue. An initial step to remove dead cells is also used. Here, we describe proof-of-concept experiments, using the biotin acceptor peptide (BAP)-low-affinity nerve growth factor receptor as the two-component protein. The first component, the BAP, can be biotinylated in specific subsets of cells expressing a particular gene by expressing the biotinylating enzyme, hBirA = humanized BirA (hBirA), under the promoter control of another gene defining the specific subpopulation. We showed that a rare population of cells (1.1% of the 13.5 days postcoital mouse embryo) could be enriched to a sufficiently high purity (84.4%). From another sample with 0.1% of our cells of interest, we achieved a 40.3% pure sample. The low cost, speed, and technical ease of the Three-step MACS also make it scalable and hence, an ideal method for preparing sufficient quantities of biological samples for sensitive, high-throughput assays.
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