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Proliferation and Differentiation of Murine Myeloid Precursor 32D/G-CSF-R Cells
Published on: February 21, 2018
[Effect of G-CSF on the proliferation and differentiation of bcr/abl(+)-CD34+ cells from CML patients]
Cheng-Yu Li1, Fan-Yi Meng, Qi-Xin Sun
1Department of Hematology, Nanfang Hospital, Nanfang Medical University, Guangzhou, China.
Insights
Granulocyte colony-stimulating factor (G-CSF) promotes the proliferation of both chronic myelocytic leukemia (CML) cells and normal CD34+ cells. While G-CSF enhances differentiation, CML cells differentiate more rapidly, independent of G-CSF.
Area of Science:
- Hematology
- Molecular Biology
- Cell Biology
Context:
- Chronic myelocytic leukemia (CML) is characterized by the bcr/abl fusion gene.
- CD34+ cells are hematopoietic stem and progenitor cells.
- Granulocyte colony-stimulating factor (G-CSF) is a cytokine that stimulates neutrophil production.
Purpose:
- To investigate the impact of G-CSF on the proliferation and differentiation of bcr/abl-positive CD34+ cells from CML patients.
- To compare the effects of G-CSF on CML-derived CD34+ cells versus normal CD34+ cells.
Summary:
- G-CSF treatment significantly increased the number of both bcr/abl(+)-CD34+ and normal CD34+ cells in a dose- and time-dependent manner.
- G-CSF induced cell-cycle progression and differentiation in both cell types, evidenced by changes in cell morphology and antigen expression (CD33, CD13).
- bcr/abl(+)-CD34+ cells exhibited more rapid differentiation compared to normal CD34+ cells, irrespective of G-CSF concentration.
Impact:
- G-CSF promotes the proliferation of both malignant and normal hematopoietic stem cells.
- Understanding G-CSF's role in CML can inform therapeutic strategies.
- The differential differentiation rates suggest distinct cellular responses in CML.
Objective:
To study the effect of granulocyte colony-stimulating factor (G-CSF) on the proliferation and differentiation of bcr/abl(+)-CD34+ cells.
Methods:
bcr/abl(+)-CD34+ cells were isolated from bone marrow of chronic myelocytic leukemia (CML) patients and were treated with 0, 10, 100, 1000 ng/ml of G-CSF for 48, 96, 144 hs. CD34 cells from normal bone marrow were used as controls. Cell proliferation was determined by trypan blue dye exclusion, cell-cycle and antigen differentiation were determined by flow cytometry and cell morphology was observed under light microscope.
Results:
The number of bcr/abl(+)-CD34+ cells was increased obviously in all groups. After cultured for 48 and 96 h, the number of bcr/abl(+)-CD34+ cells at G-CSF 10 ng/ml group was significantly higher than that in G-CSF 0 ng/ml group (P < 0.05) , the number of normal CD34 cells was increased only in the presence of G-CSF. After cultured for 48, 96 and 144 h, the cell number in G-CSF 100 ng/ml group was significantly higher than that in G-CSF 0 ng/ml group (P < 0.05, P < 0.01, P < 0.01, respectively). After cultured for 144 h, the cell percentages in G0/G1 phase for bcr/abl(+)-CD34+ cells in G-CSF 10, 100, 1000 ng/ml groups were significantly less than that in G-CSF 0 ng/ml group (P < 0. 05), and that for normal CD34 cells in G-CSF 10, 100, 1000 ng/ml groups were significantly less than that of G-CSF 0 ng/ml group after cultured for 48 and 96 h. The expressions of CD34 on bcr/abl(+)-CD34+ cells and normal CD34+ cells were decreased along with the culture duration, accompanied by the expression of CD33 and CD13 increased first and decreased later, which was not correlated with the concentration of G-CSF. Both bcr/abl(+)-CD34+ cells and normal CD34+ cells showed mature morphology along with proliferation and differentiation.
Conclusions:
G-CSF promotes proliferation of both bcr/abl(+)-CD34+ cells and normal CD34+ cells, but not necessary for the former, and the former differentiates more rapidly than the latter does, but both was independent of G-CSF.
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