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- W2078697135 abstract "Abstract Detailed studies on the biology of Langerhans cells (LC), which account for only 1-3% of all epidermal cells, require isolation from their cutaneous symbionts. Several techniques of LC isolation have been reported, including positive enrichment with mAb coupled to immuno-magnetic beads. The disadvantage of this technique is the size of the beads (= 2-5 μm), which can interfere with subsequent phenotypic and functional analyses. This limitation prompted us to test wheter paramagnetic microbeads (15 nm) employed by the MACS™ system could be used to purify LC from human skin. To isolate fresh LC (fLC), epidermal cell suspensions (EC) were stained with anti-CD la mAb and with appropriate secondary reagents conjugated to microbeads and to FITC. They were then passed over a separation column and exposed to a strong magnetic field. Thereafter both CD la-depleted and CD la-enriched cells were collected. Cultured LC (cLC) were isolated by staining 72-h cultured EC with anti-HLA-DR mAb followed by the same isolation procedure. Using this technique, we could routinely isolate viable EC that were 45-88% CD la or HLA-DR+ as determined by FACS. Two-color FACS analysis demonstrated the majority of MACS-purified cells to be CDla+/HLA-DR+, indicating that they were indeed LC. By transmission electron microscopy (TEM), the MACS-purified CDla+/HLA-DR+ cells showed typical ultrastructural characteristics of LC. Furthermore, MACS-purified fLC or cLC were functionally intact, because they stimulated the proliferation of alloreactive T cells in a primary, one-way, mixed epidermal cell leukocyte reaction (MECLR). We conclude that MACS-separation is an efficient and rapid method to isolate human fLC and cLC of high purity and unimpaired function." @default.
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- W2078697135 date "1995-06-01" @default.
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- W2078697135 title "Rapid purification of human Langerhans cells using paramagnetic microbeads" @default.
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- W2078697135 doi "https://doi.org/10.1111/j.1600-0625.1995.tb00239.x" @default.
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