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Stem cell factor amplifies newborn and sickle erythropoiesis in liquid cultures
R S Weinberg1, J C Thomson, R Lao
1Polly Annenberg Levee Hematology Center, Department of Medicine, Mount Sinai School of Medicine, New York, NY 10029.
Insights
Stem cell factor (SCF) significantly amplifies erythroid progenitor cells in a two-phase culture system. SCF, combined with erythropoietin (Ep), enhances erythroblast and erythrocyte differentiation, particularly in sickle cell anemia and newborn samples.
Area of Science:
- Hematology
- Cell Biology
- Stem Cell Research
Background:
- Erythroblast differentiation is crucial for red blood cell production.
- Understanding progenitor cell expansion is key for regenerative medicine.
- Sickle cell anemia presents unique challenges in erythropoiesis.
Purpose of the Study:
- To investigate the amplification and differentiation of erythroblasts using a two-phase liquid-culture system.
- To evaluate the effects of stem cell factor (SCF) and erythropoietin (Ep) on erythroid progenitor cells.
- To compare the responses of cells from normal adults, newborns, and sickle cell anemia patients.
Main Methods:
- A two-phase liquid-culture system was employed, starting with mononuclear cells.
- Phase 1 involved culture in medium with fetal bovine serum (FBS), SCF, or conditioned medium (CM).
- Phase 2 utilized erythropoietin (Ep) with or without SCF/CM, or methylcellulose cultures.
Main Results:
- SCF in phase 1 increased late progenitors (CFU-E), with higher numbers in sickle and newborn samples.
- Phase 2 cultures with SCF and Ep showed significant cell number increases: 30-fold (sickle), 24-fold (newborn), and 4-fold (adult).
- SCF's effects were most pronounced in sickle cell anemia and newborn cultures.
Conclusions:
- Erythroid progenitor cells can survive for at least 7 days without erythropoietin (Ep) in the presence of FBS.
- Stem cell factor (SCF) effectively amplifies erythroid progenitor cells.
- SCF combined with Ep enhances the final number of differentiated erythroid cells, with notable efficacy in sickle cell and newborn samples.
Abstract:
A two-phase liquid-culture system was used to substantially amplify and differentiate erythroblasts, starting with mononuclear cells from the blood of normal adults, newborn infants, and patients with sickle cell anemia. After the first 7 days (phase 1), in medium plus fetal bovine serum (FBS) alone, or in combination with stem cell factor (SCF) or conditioned medium (CM), the cell number was unchanged, and the cells all looked like lymphocytes. These cells were then diluted into medium with erythropoietin (Ep) alone, with Ep and either SCF or CM, or in methylcellulose with the same factors (phase 2). After 14 days in liquid phase 2 with SCF and Ep, the cell numbers increased an average of 30-fold in the sickle, 24-fold in the newborn, and 4-fold in the normal adult cultures; almost all the cells were erythroblasts and erythrocytes. SCF in phase 1 increased the number of late progenitors (CFU-E) assayed in methylcellulose, with the largest number in sickle, followed by newborn cultures and then adult cultures. We conclude that erythroid progenitor cells survive for at least 7 days without Ep (but with FBS). Progenitor cells are amplified, particularly with SCF. Later in culture, SCF with Ep increases the final number of differentiated erythroid cells. Both the early and the late effects of SCF are most effective in sickle, followed by newborn cultures and then adult cultures.