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Purified unfractionated G‐CSF/chemotherapy mobilized CD34 + peripheral blood progenitors and not bone marrow CD34 + progenitors undergo selective erythroid differentiation in liquid culture in the presence of erythropoietin and stem cell factor
Author(s) -
PIERELLI Luca,
Scambia Giovanni,
Menichella Giacomo,
Fattorossi Andrea,
Ciarli Marina,
Bonanno Giuseppina,
Battaglia Alessandra,
D'Onofrio Giuseppe,
Benedetti Panici Pierluigi,
Iacone Antonio,
Mancuso Salvatore,
Leone Giuseppe
Publication year - 1997
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.1997.8632491.x
Subject(s) - cd34 , clonogenic assay , bone marrow , progenitor cell , erythropoietin , stem cell factor , haematopoiesis , stem cell , biology , immunology , interleukin 3 , cytokine , cellular differentiation , microbiology and biotechnology , cell culture , chemistry , endocrinology , antigen , biochemistry , interleukin 21 , genetics , gene , cd8
A combination of erythropoietin (EPO) plus stem cell factor (SCF) drove purified unfractionated granulocyte colony stimulating factor (G‐CSF)/chemotherapy mobilized peripheral blood CD34 + cells to selective erythroid differentiation in liquid culture with an average 28‐fold increase in the total cell number after 21 d. From day 6 of culture, cytologic and cytofluorimetric characterization revealed that cultured cells belonged to the erythroid lineage with a gradual wave of maturation along the erythroid pathway to terminal cells. A similar pattern of erythroid differentiation was observed when the same peripheral blood CD34 + cells were cultured with EPO plus SCF in serum‐free medium. This cytokine combination produced selective erythroid differentiation with the complete exhaustion of the clonogenic potential on day 21. In parallel experiments the same circulating CD34 + cells underwent granulocytic/monocytic differentiation in liquid culture in response to granulocyte‐macrophage colony stimulating factor (GM‐CSF), interleukin‐3 (IL‐3) and SCF, demonstrating that these CD34 + progenitors had intact pluripotent differentiating potential. Conversely, bone marrow CD34 + cells isolated from bone marrow allografts were unable to selectively differentiate along the erythroid pathway when they were exposed to EPO plus SCF combination. However, these cells maintained a greater number of colony forming cells on day 21 of culture compared to mobilized peripheral blood CD34 + cells. This model is a simple and reliable way to obtain selective erythroid differentiation of peripheral blood G‐CSF/chemotherapy mobilized CD34 + progenitor cells in liquid culture. The absence of cytokines such as GM‐CSF and IL‐3 in the culture medium permits studies on in vitro erythropoiesis without disturbance of prevalent myelopoiesis.