Cytokine-dependent ex vivo expansion of early subsets of CD34+ cord blood myeloid progenitors is enhanced by cord blood plasma, but expansion of the more mature subsets of progenitors is favored

L. Ruggieri, S. Heimfeld, Hal Broxmeyer

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Abstract

Expansion of stem/progenitor cells has important implications for transplantation. We recently reported that a factor or factors in cord blood (CB), but not adult peripheral blood (PB), plasma enhanced replating of granulocyte erythroid macrophage megakaryocyte colony-forming units (CFU-GEMM) progenitors, a measure of self-renewal capacity. In this context, we evaluated effects of CB plasma, in comparison with PB plasma and fetal bovine serum (FBS), on ex vivo expansion of CD34+ column-separated (72-98% CD34+) CB cells using stroma-free cultures in the absence and presence of either PIXY321 (a granulocyte-macrophage colony-stimulating factor/interleukin-3 [GM-CSF/IL-3] fusion protein), IL-3 + IL-6 + IL-1, or steel factor (SLF) -/+ PIXY. CB plasma, PB plasma, or FBS alone did not sustain cell numbers. Combinations of CB plasma + SLF + PIXY induced maximal cumulative nucleated cell expansion (1044-fold), which was greater than that of PB plasma plus cytokines (633-fold) and FBS plus cytokines (142-fold). Total CD34+ cells peaked by day 7 with 7-fold expansion in the presence of CB plasma + SLF + PIXY compared with PB plasma or FBS with these same cytokines (threefold each). By day 7, total CFU-GEMM production in the presence of either PIXY, SLF + PIXY, or IL-3 + IL-6 + IL-1 was greater with CB plasma (maximum 11.4-fold average increases) than with PB plasma (6.8-fold increase). These increases were greater than with FBS. However, PB plasma was at least as good as CB plasma for expansion of immature and mature subsets of CFU-GM. The frequency of progenitors decreased with time, and expansion was coupled with differentiation. Although the proliferative capacity of CFU-GEMM was maintained, the capacity of CFU-GEMM to be replated decreased after time in suspension culture, suggesting age-related commitment of cells. Moreover, with plasma + SLE + PIXY for 7 days, expansion of more mature CFU-GM (responsive to GM-CSF) was greater (16-146-fold with CB plasma and 31-208-fold with PB plasma) than immature CFU-GM (responsive to GM-CSF + SFU) (4- to 14-fold with CB plasma and 6- to 17-fold with PB plasma). The results suggest that CB plasma enhances expansion of CFU-GEMM to a greater extent than PB plasma or FBS, but expansion in these cultures favors more mature subsets of cells.

Original languageEnglish
Pages (from-to)436-454
Number of pages19
JournalBlood Cells
Volume20
Issue number2-3
StatePublished - 1994

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Fetal Blood
Cytokines
Myeloid Progenitor Cells
Stem Cell Factor
Interleukin-3
Granulocyte-Macrophage Colony-Stimulating Factor
Granulocyte-Macrophage Progenitor Cells
Serum
Interleukin-1
Interleukin-6
Stem Cells

Keywords

  • cord blood
  • cord blood plasma
  • cytokines
  • expansion
  • stem/progenitor cells

ASJC Scopus subject areas

  • Hematology

Cite this

@article{1f34af0537db412fa5338e128a46ca97,
title = "Cytokine-dependent ex vivo expansion of early subsets of CD34+ cord blood myeloid progenitors is enhanced by cord blood plasma, but expansion of the more mature subsets of progenitors is favored",
abstract = "Expansion of stem/progenitor cells has important implications for transplantation. We recently reported that a factor or factors in cord blood (CB), but not adult peripheral blood (PB), plasma enhanced replating of granulocyte erythroid macrophage megakaryocyte colony-forming units (CFU-GEMM) progenitors, a measure of self-renewal capacity. In this context, we evaluated effects of CB plasma, in comparison with PB plasma and fetal bovine serum (FBS), on ex vivo expansion of CD34+ column-separated (72-98{\%} CD34+) CB cells using stroma-free cultures in the absence and presence of either PIXY321 (a granulocyte-macrophage colony-stimulating factor/interleukin-3 [GM-CSF/IL-3] fusion protein), IL-3 + IL-6 + IL-1, or steel factor (SLF) -/+ PIXY. CB plasma, PB plasma, or FBS alone did not sustain cell numbers. Combinations of CB plasma + SLF + PIXY induced maximal cumulative nucleated cell expansion (1044-fold), which was greater than that of PB plasma plus cytokines (633-fold) and FBS plus cytokines (142-fold). Total CD34+ cells peaked by day 7 with 7-fold expansion in the presence of CB plasma + SLF + PIXY compared with PB plasma or FBS with these same cytokines (threefold each). By day 7, total CFU-GEMM production in the presence of either PIXY, SLF + PIXY, or IL-3 + IL-6 + IL-1 was greater with CB plasma (maximum 11.4-fold average increases) than with PB plasma (6.8-fold increase). These increases were greater than with FBS. However, PB plasma was at least as good as CB plasma for expansion of immature and mature subsets of CFU-GM. The frequency of progenitors decreased with time, and expansion was coupled with differentiation. Although the proliferative capacity of CFU-GEMM was maintained, the capacity of CFU-GEMM to be replated decreased after time in suspension culture, suggesting age-related commitment of cells. Moreover, with plasma + SLE + PIXY for 7 days, expansion of more mature CFU-GM (responsive to GM-CSF) was greater (16-146-fold with CB plasma and 31-208-fold with PB plasma) than immature CFU-GM (responsive to GM-CSF + SFU) (4- to 14-fold with CB plasma and 6- to 17-fold with PB plasma). The results suggest that CB plasma enhances expansion of CFU-GEMM to a greater extent than PB plasma or FBS, but expansion in these cultures favors more mature subsets of cells.",
keywords = "cord blood, cord blood plasma, cytokines, expansion, stem/progenitor cells",
author = "L. Ruggieri and S. Heimfeld and Hal Broxmeyer",
year = "1994",
language = "English",
volume = "20",
pages = "436--454",
journal = "Blood Cells, Molecules, and Diseases",
issn = "1079-9796",
publisher = "Academic Press Inc.",
number = "2-3",

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TY - JOUR

T1 - Cytokine-dependent ex vivo expansion of early subsets of CD34+ cord blood myeloid progenitors is enhanced by cord blood plasma, but expansion of the more mature subsets of progenitors is favored

AU - Ruggieri, L.

AU - Heimfeld, S.

AU - Broxmeyer, Hal

PY - 1994

Y1 - 1994

N2 - Expansion of stem/progenitor cells has important implications for transplantation. We recently reported that a factor or factors in cord blood (CB), but not adult peripheral blood (PB), plasma enhanced replating of granulocyte erythroid macrophage megakaryocyte colony-forming units (CFU-GEMM) progenitors, a measure of self-renewal capacity. In this context, we evaluated effects of CB plasma, in comparison with PB plasma and fetal bovine serum (FBS), on ex vivo expansion of CD34+ column-separated (72-98% CD34+) CB cells using stroma-free cultures in the absence and presence of either PIXY321 (a granulocyte-macrophage colony-stimulating factor/interleukin-3 [GM-CSF/IL-3] fusion protein), IL-3 + IL-6 + IL-1, or steel factor (SLF) -/+ PIXY. CB plasma, PB plasma, or FBS alone did not sustain cell numbers. Combinations of CB plasma + SLF + PIXY induced maximal cumulative nucleated cell expansion (1044-fold), which was greater than that of PB plasma plus cytokines (633-fold) and FBS plus cytokines (142-fold). Total CD34+ cells peaked by day 7 with 7-fold expansion in the presence of CB plasma + SLF + PIXY compared with PB plasma or FBS with these same cytokines (threefold each). By day 7, total CFU-GEMM production in the presence of either PIXY, SLF + PIXY, or IL-3 + IL-6 + IL-1 was greater with CB plasma (maximum 11.4-fold average increases) than with PB plasma (6.8-fold increase). These increases were greater than with FBS. However, PB plasma was at least as good as CB plasma for expansion of immature and mature subsets of CFU-GM. The frequency of progenitors decreased with time, and expansion was coupled with differentiation. Although the proliferative capacity of CFU-GEMM was maintained, the capacity of CFU-GEMM to be replated decreased after time in suspension culture, suggesting age-related commitment of cells. Moreover, with plasma + SLE + PIXY for 7 days, expansion of more mature CFU-GM (responsive to GM-CSF) was greater (16-146-fold with CB plasma and 31-208-fold with PB plasma) than immature CFU-GM (responsive to GM-CSF + SFU) (4- to 14-fold with CB plasma and 6- to 17-fold with PB plasma). The results suggest that CB plasma enhances expansion of CFU-GEMM to a greater extent than PB plasma or FBS, but expansion in these cultures favors more mature subsets of cells.

AB - Expansion of stem/progenitor cells has important implications for transplantation. We recently reported that a factor or factors in cord blood (CB), but not adult peripheral blood (PB), plasma enhanced replating of granulocyte erythroid macrophage megakaryocyte colony-forming units (CFU-GEMM) progenitors, a measure of self-renewal capacity. In this context, we evaluated effects of CB plasma, in comparison with PB plasma and fetal bovine serum (FBS), on ex vivo expansion of CD34+ column-separated (72-98% CD34+) CB cells using stroma-free cultures in the absence and presence of either PIXY321 (a granulocyte-macrophage colony-stimulating factor/interleukin-3 [GM-CSF/IL-3] fusion protein), IL-3 + IL-6 + IL-1, or steel factor (SLF) -/+ PIXY. CB plasma, PB plasma, or FBS alone did not sustain cell numbers. Combinations of CB plasma + SLF + PIXY induced maximal cumulative nucleated cell expansion (1044-fold), which was greater than that of PB plasma plus cytokines (633-fold) and FBS plus cytokines (142-fold). Total CD34+ cells peaked by day 7 with 7-fold expansion in the presence of CB plasma + SLF + PIXY compared with PB plasma or FBS with these same cytokines (threefold each). By day 7, total CFU-GEMM production in the presence of either PIXY, SLF + PIXY, or IL-3 + IL-6 + IL-1 was greater with CB plasma (maximum 11.4-fold average increases) than with PB plasma (6.8-fold increase). These increases were greater than with FBS. However, PB plasma was at least as good as CB plasma for expansion of immature and mature subsets of CFU-GM. The frequency of progenitors decreased with time, and expansion was coupled with differentiation. Although the proliferative capacity of CFU-GEMM was maintained, the capacity of CFU-GEMM to be replated decreased after time in suspension culture, suggesting age-related commitment of cells. Moreover, with plasma + SLE + PIXY for 7 days, expansion of more mature CFU-GM (responsive to GM-CSF) was greater (16-146-fold with CB plasma and 31-208-fold with PB plasma) than immature CFU-GM (responsive to GM-CSF + SFU) (4- to 14-fold with CB plasma and 6- to 17-fold with PB plasma). The results suggest that CB plasma enhances expansion of CFU-GEMM to a greater extent than PB plasma or FBS, but expansion in these cultures favors more mature subsets of cells.

KW - cord blood

KW - cord blood plasma

KW - cytokines

KW - expansion

KW - stem/progenitor cells

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JO - Blood Cells, Molecules, and Diseases

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SN - 1079-9796

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