Improved FACS analysis confirms generation of immature dendritic cells in long-term stromal-dependent spleen cultures

KP Ni, HC O'Neill

Research output: Contribution to journalArticleResearchpeer-review

15 Citations (Scopus)

Abstract

Cells produced in spleen stroma-dependent long-term cultures (LTC) have now been clearly defined as dendritic cells (DC). Characterization of cells by antibody staining and FAGS analysis has only been possible using a procedure to quench the high autofluorescence of DC produced in LTC (LTC-DC). The population of large cells produced by the established LTC-X1 culture are homogeneously positive for a number of cell-surface markers expressed by DC. These include CD11c, CD11b, Dec-205, Fc receptor and CD86. They also express markers detectable with the F4/80 and 33D1 antibodies. Cells produced in LTC do not uniformly express the MHC II marker, consistent with an immature DC phenotype. Most cells are weakly positive for MMC II with a small subset of highly positive cells. The quenching method involves staining cells with crystal violet dye, which is taken up within the cell. The importance of optimizing fluorescent antibody staining assays for delineating DC subsets is indicated and the LTC system is established as a valuable and continuous source of DC precursors.

Original languageEnglish
Pages (from-to)196-204
Number of pages9
JournalImmunology and Cell Biology
Volume78
Issue number3
DOIs
Publication statusPublished - Jun 2000
Externally publishedYes

Cite this

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title = "Improved FACS analysis confirms generation of immature dendritic cells in long-term stromal-dependent spleen cultures",
abstract = "Cells produced in spleen stroma-dependent long-term cultures (LTC) have now been clearly defined as dendritic cells (DC). Characterization of cells by antibody staining and FAGS analysis has only been possible using a procedure to quench the high autofluorescence of DC produced in LTC (LTC-DC). The population of large cells produced by the established LTC-X1 culture are homogeneously positive for a number of cell-surface markers expressed by DC. These include CD11c, CD11b, Dec-205, Fc receptor and CD86. They also express markers detectable with the F4/80 and 33D1 antibodies. Cells produced in LTC do not uniformly express the MHC II marker, consistent with an immature DC phenotype. Most cells are weakly positive for MMC II with a small subset of highly positive cells. The quenching method involves staining cells with crystal violet dye, which is taken up within the cell. The importance of optimizing fluorescent antibody staining assays for delineating DC subsets is indicated and the LTC system is established as a valuable and continuous source of DC precursors.",
author = "KP Ni and HC O'Neill",
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Improved FACS analysis confirms generation of immature dendritic cells in long-term stromal-dependent spleen cultures. / Ni, KP; O'Neill, HC.

In: Immunology and Cell Biology, Vol. 78, No. 3, 06.2000, p. 196-204.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Improved FACS analysis confirms generation of immature dendritic cells in long-term stromal-dependent spleen cultures

AU - Ni, KP

AU - O'Neill, HC

PY - 2000/6

Y1 - 2000/6

N2 - Cells produced in spleen stroma-dependent long-term cultures (LTC) have now been clearly defined as dendritic cells (DC). Characterization of cells by antibody staining and FAGS analysis has only been possible using a procedure to quench the high autofluorescence of DC produced in LTC (LTC-DC). The population of large cells produced by the established LTC-X1 culture are homogeneously positive for a number of cell-surface markers expressed by DC. These include CD11c, CD11b, Dec-205, Fc receptor and CD86. They also express markers detectable with the F4/80 and 33D1 antibodies. Cells produced in LTC do not uniformly express the MHC II marker, consistent with an immature DC phenotype. Most cells are weakly positive for MMC II with a small subset of highly positive cells. The quenching method involves staining cells with crystal violet dye, which is taken up within the cell. The importance of optimizing fluorescent antibody staining assays for delineating DC subsets is indicated and the LTC system is established as a valuable and continuous source of DC precursors.

AB - Cells produced in spleen stroma-dependent long-term cultures (LTC) have now been clearly defined as dendritic cells (DC). Characterization of cells by antibody staining and FAGS analysis has only been possible using a procedure to quench the high autofluorescence of DC produced in LTC (LTC-DC). The population of large cells produced by the established LTC-X1 culture are homogeneously positive for a number of cell-surface markers expressed by DC. These include CD11c, CD11b, Dec-205, Fc receptor and CD86. They also express markers detectable with the F4/80 and 33D1 antibodies. Cells produced in LTC do not uniformly express the MHC II marker, consistent with an immature DC phenotype. Most cells are weakly positive for MMC II with a small subset of highly positive cells. The quenching method involves staining cells with crystal violet dye, which is taken up within the cell. The importance of optimizing fluorescent antibody staining assays for delineating DC subsets is indicated and the LTC system is established as a valuable and continuous source of DC precursors.

U2 - 10.1046/j.1440-1711.2000.00897.x

DO - 10.1046/j.1440-1711.2000.00897.x

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EP - 204

JO - Australian Journal of Experimental Biology and Medical Science

JF - Australian Journal of Experimental Biology and Medical Science

SN - 0818-9641

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ER -