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Titolo:
Establishment of serum-free pre-colony forming unit assays for differentiation of primitive hematopoietic progenitors: serum induces early macrophagedifferentiation and inhibits early erythroid differentiation of CD34(++)CD38(-) cells
Autore:
Willems, R; Henckaerts, E; Lenjou, M; Nijs, G; Rodrigus, I; Moulijn, AC; Slegers, H; Berneman, ZN; Van Bockstaele, DR;
Indirizzi:
Univ Antwerp, Antwerp Univ Hosp, Lab Expt Hematol, UIA UZA, B-2650 Edegem,Belgium Univ Antwerp Edegem Belgium B-2650 matol, UIA UZA, B-2650 Edegem,Belgium Univ Instelling Antwerp, Lab Cellular Biochem, B-2610 Wilrijk, Belgium Univ Instelling Antwerp Wilrijk Belgium B-2610 , B-2610 Wilrijk, Belgium UZA, UIA, Antwerp Univ Hosp, Dept Cardiac Surg, B-2650 Edegem, Belgium UZA Edegem Belgium B-2650 osp, Dept Cardiac Surg, B-2650 Edegem, Belgium
Titolo Testata:
ANNALS OF HEMATOLOGY
fascicolo: 1, volume: 80, anno: 2001,
pagine: 17 - 25
SICI:
0939-5555(200101)80:1<17:EOSPFU>2.0.ZU;2-6
Fonte:
ISI
Lingua:
ENG
Soggetto:
HUMAN BONE-MARROW; EX-VIVO EXPANSION; FLT3 LIGAND; STEM-CELLS; CORD-BLOOD; C-KIT; MONOCYTE DIFFERENTIATION; CD34(+)CD38(-) CELLS; STROMAL CELLS; IN-VITRO;
Keywords:
hematopoietic progenitor; Flt3/Flk2 ligand; serum-free medium; monocyte/macrophage differentiation; erythroid differentiation;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
39
Recensione:
Indirizzi per estratti:
Indirizzo: Berneman, ZN Univ Antwerp, Antwerp Univ Hosp, Lab Expt Hematol, UIA UZA, Wilrijkstr 10,B-2650 Edegem, Belgium Univ Antwerp Wilrijkstr 10 Edegem Belgium B-2650 em, Belgium
Citazione:
R. Willems et al., "Establishment of serum-free pre-colony forming unit assays for differentiation of primitive hematopoietic progenitors: serum induces early macrophagedifferentiation and inhibits early erythroid differentiation of CD34(++)CD38(-) cells", ANN HEMATOL, 80(1), 2001, pp. 17-25

Abstract

In this report we show that serum has differentiation-inducing effects on primitive hematopoietic progenitor cells with the CD34(++)CD38(-) immunophenotype. Using the pre-colony forming unit (pre-CFU) assay as a model for early myelopoiesis, we compared the effects of serum-containing and serum-free media and evaluated different cytokine cocktails [interleukin (IL)-1, IL-3, IL-6, kit ligand with and without the Flt3/ Flk2 ligand (FL)]. In this assay, pre-CFUs are defined as cells unable to form colonies when plated directly in semi-solid assays, but which can differentiate into CFUs when cultured in liquid medium containing early-acting, cytokines. In one of the investigated serum-free media, the average myeloid expansion in liquid medium reached up to more than 50% of that obtained in serum-containing medium. Inaddition, our experiments revealed differences in the clonogenic output between cells cultured in serum-free medium and those cultured in serum-containing, medium, demonstrating that serum has a monocyte differentiation-inducing effect on primitive hematopoietic progenitors. Also in serum-free medium, higher proportions of erythroid progenitors were generated. These differentiation-inducing effects of serum further emphasize the need for serum-free culture protocols for hematopoietic graft engineering. Addition of FL to the culture media ameliorated cellular expansion and resulted in a decrease in the proportion of erythroid and granulocyte progenitors and an increase in the proportion of monocyte progenitors. In conclusion, this study shows that good serum-free conditions are available for differentiation assayswith primitive hematopoietic progenitors and demonstrates that serum and FL have biasing effects on the initial phase of hematopoietic differentiation, favoring the monocyte lineage.

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Documento generato il 04/04/20 alle ore 21:01:59