Long-term generation of human mast cells in serum-free cultures of CD34+ cord blood cells stimulated with stem cell factor and interleukin-3

Brigitte Durand, Giovanni Migliaccio, Nelson Shu-Sang Yee, Keith Eddleman, Tellervo Huima-Byron, Anna Rita Migliaccio, John W. Adamson

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

The generation of murine mast cells is supported by several cytokines, and mast cell lines are frequently established in long-term cultures of normal murine marrow cells. In contrast, growth of human mast cells was initially dependent on coculture with murine fibroblasts. The growth factor produced by murine fibroblasts and required to observe differentiation of human mast cells is attributable in part to stem cell factor (SCF). However, other factors are likely involved. We have previously shown that the combination of SCF and interleukin-3 (IL-3) efficiently sustains proliferation and differentiation of colony-forming cells (CFCs) from pre-CFC enriched from human umbilical cord blood by CD34+ selection. With periodic medium changes and the addition of fresh growth factors, five consecutive cultures of different cord blood samples gave rise to differentiated cells and CFCs for more than 2 months. Although differentiated cells continued to be generated for more than 5 months, CFCs were no longer detectable by day 50 of culture. The cells have the morphology of immature mast cells, are Toluidine blue positive, are karyotypically normal, are CD33+, CD34-, CD45+, c-kit-, and c-fms-, and die in the absence of either SCF or IL-3. These cells do not form colonies in semisolid culture and are propagated in liquid culture stimulated with SCF and IL-3 at a seeding concentration of no less than 104 cells/mL. At refeedings, the cultures contain a high number (>50%) of dead cells and have a doubling time ranging from 5 to 12 days. This suggests that subsets of the cell population die because of a requirement for a growth factor other than SCF or IL-3. These results indicate that the combination of cord blood progenitor and stem cells, plus a cocktail of growth factors including SCF and IL-3, is capable with high efficiency of giving rise in serum-deprived culture to human mast cells that behave like factor-dependent cell lines. These cells may represent a useful tool for studies of human mast cell differentiation and leukemia.

Original languageEnglish (US)
Pages (from-to)3667-3674
Number of pages8
JournalBlood
Volume84
Issue number11
StatePublished - Dec 1 1994

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Stem Cell Factor
Interleukin-3
Fetal Blood
Cell culture
Mast Cells
Blood Cells
Blood
Cells
Intercellular Signaling Peptides and Proteins
Serum
Fibroblasts
Tolonium Chloride
Stem cells
Mast-Cell Leukemia
Stem Cells
Cytokines
Cell Line
Liquids
Coculture Techniques
Cell Differentiation

All Science Journal Classification (ASJC) codes

  • Hematology

Cite this

Durand, B., Migliaccio, G., Yee, N. S-S., Eddleman, K., Huima-Byron, T., Migliaccio, A. R., & Adamson, J. W. (1994). Long-term generation of human mast cells in serum-free cultures of CD34+ cord blood cells stimulated with stem cell factor and interleukin-3. Blood, 84(11), 3667-3674.
Durand, Brigitte ; Migliaccio, Giovanni ; Yee, Nelson Shu-Sang ; Eddleman, Keith ; Huima-Byron, Tellervo ; Migliaccio, Anna Rita ; Adamson, John W. / Long-term generation of human mast cells in serum-free cultures of CD34+ cord blood cells stimulated with stem cell factor and interleukin-3. In: Blood. 1994 ; Vol. 84, No. 11. pp. 3667-3674.
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abstract = "The generation of murine mast cells is supported by several cytokines, and mast cell lines are frequently established in long-term cultures of normal murine marrow cells. In contrast, growth of human mast cells was initially dependent on coculture with murine fibroblasts. The growth factor produced by murine fibroblasts and required to observe differentiation of human mast cells is attributable in part to stem cell factor (SCF). However, other factors are likely involved. We have previously shown that the combination of SCF and interleukin-3 (IL-3) efficiently sustains proliferation and differentiation of colony-forming cells (CFCs) from pre-CFC enriched from human umbilical cord blood by CD34+ selection. With periodic medium changes and the addition of fresh growth factors, five consecutive cultures of different cord blood samples gave rise to differentiated cells and CFCs for more than 2 months. Although differentiated cells continued to be generated for more than 5 months, CFCs were no longer detectable by day 50 of culture. The cells have the morphology of immature mast cells, are Toluidine blue positive, are karyotypically normal, are CD33+, CD34-, CD45+, c-kit-, and c-fms-, and die in the absence of either SCF or IL-3. These cells do not form colonies in semisolid culture and are propagated in liquid culture stimulated with SCF and IL-3 at a seeding concentration of no less than 104 cells/mL. At refeedings, the cultures contain a high number (>50{\%}) of dead cells and have a doubling time ranging from 5 to 12 days. This suggests that subsets of the cell population die because of a requirement for a growth factor other than SCF or IL-3. These results indicate that the combination of cord blood progenitor and stem cells, plus a cocktail of growth factors including SCF and IL-3, is capable with high efficiency of giving rise in serum-deprived culture to human mast cells that behave like factor-dependent cell lines. These cells may represent a useful tool for studies of human mast cell differentiation and leukemia.",
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Durand, B, Migliaccio, G, Yee, NS-S, Eddleman, K, Huima-Byron, T, Migliaccio, AR & Adamson, JW 1994, 'Long-term generation of human mast cells in serum-free cultures of CD34+ cord blood cells stimulated with stem cell factor and interleukin-3', Blood, vol. 84, no. 11, pp. 3667-3674.

Long-term generation of human mast cells in serum-free cultures of CD34+ cord blood cells stimulated with stem cell factor and interleukin-3. / Durand, Brigitte; Migliaccio, Giovanni; Yee, Nelson Shu-Sang; Eddleman, Keith; Huima-Byron, Tellervo; Migliaccio, Anna Rita; Adamson, John W.

In: Blood, Vol. 84, No. 11, 01.12.1994, p. 3667-3674.

Research output: Contribution to journalArticle

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AU - Durand, Brigitte

AU - Migliaccio, Giovanni

AU - Yee, Nelson Shu-Sang

AU - Eddleman, Keith

AU - Huima-Byron, Tellervo

AU - Migliaccio, Anna Rita

AU - Adamson, John W.

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Y1 - 1994/12/1

N2 - The generation of murine mast cells is supported by several cytokines, and mast cell lines are frequently established in long-term cultures of normal murine marrow cells. In contrast, growth of human mast cells was initially dependent on coculture with murine fibroblasts. The growth factor produced by murine fibroblasts and required to observe differentiation of human mast cells is attributable in part to stem cell factor (SCF). However, other factors are likely involved. We have previously shown that the combination of SCF and interleukin-3 (IL-3) efficiently sustains proliferation and differentiation of colony-forming cells (CFCs) from pre-CFC enriched from human umbilical cord blood by CD34+ selection. With periodic medium changes and the addition of fresh growth factors, five consecutive cultures of different cord blood samples gave rise to differentiated cells and CFCs for more than 2 months. Although differentiated cells continued to be generated for more than 5 months, CFCs were no longer detectable by day 50 of culture. The cells have the morphology of immature mast cells, are Toluidine blue positive, are karyotypically normal, are CD33+, CD34-, CD45+, c-kit-, and c-fms-, and die in the absence of either SCF or IL-3. These cells do not form colonies in semisolid culture and are propagated in liquid culture stimulated with SCF and IL-3 at a seeding concentration of no less than 104 cells/mL. At refeedings, the cultures contain a high number (>50%) of dead cells and have a doubling time ranging from 5 to 12 days. This suggests that subsets of the cell population die because of a requirement for a growth factor other than SCF or IL-3. These results indicate that the combination of cord blood progenitor and stem cells, plus a cocktail of growth factors including SCF and IL-3, is capable with high efficiency of giving rise in serum-deprived culture to human mast cells that behave like factor-dependent cell lines. These cells may represent a useful tool for studies of human mast cell differentiation and leukemia.

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Durand B, Migliaccio G, Yee NS-S, Eddleman K, Huima-Byron T, Migliaccio AR et al. Long-term generation of human mast cells in serum-free cultures of CD34+ cord blood cells stimulated with stem cell factor and interleukin-3. Blood. 1994 Dec 1;84(11):3667-3674.