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Under HEMA conditions, self-replication of human erythroblasts is limited by autophagic death.
- Source :
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Blood cells, molecules & diseases [Blood Cells Mol Dis] 2011 Oct 15; Vol. 47 (3), pp. 182-97. Date of Electronic Publication: 2011 Jul 20. - Publication Year :
- 2011
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Abstract
- The number of erythroblasts generated ex-vivo under human-erythroid massive-amplification conditions by mononuclear cells from one unit of adult blood (~10(10)) are insufficient for transfusion (~10(12) red cells), emphasizing the need for studies to characterize cellular interactions during culture to increase erythroblast production. To identify the cell populations which generate erythroblasts under human-erythroid-massive-amplification conditions and the factors that limit proliferation, day 10 non-erythroblasts and immature- and mature-erythroblasts were separated by sorting, labelled with carboxyfluorescein-diacetate-succinimidyl-ester and re-cultured either under these conditions (for proliferation, maturation and/or apoptosis/autophagy determinations) or in semisolid media (for progenitor cell determination). Non-erythroblasts contained 54% of the progenitor cells but did not grow under human-erythroid-massive-amplification conditions. Immature-erythroblasts contained 25% of the progenitor cells and generated erythroblasts under human-erythroid-massive-amplification conditions (FI at 48 h=2.57±1.15). Mature-erythroblasts did not generate colonies and died in human-erythroid-massive-amplification conditions. In sequential sorting/re-culture experiments, immature-erythroblasts retained the ability to generate erythroblasts for 6 days and generated 2-5-fold more cells than the corresponding unfractionated population, suggesting that mature-erythroblasts may limit erythroblast expansion. In co-cultures of carboxyfluorescein-diacetate-succinimidyl-ester-labelled-immature-erythroblasts with mature-erythroblasts at increasing ratios, cell numbers did not increase and proliferation, maturation and apoptotic rates were unchanged. However, Acridine Orange staining (a marker for autophagic death) increased from ~3.2% in cultures with immature-erythroblasts alone to 14-22% in cultures of mature-erythroblasts with and without immature-erythroblasts. In conclusion, these data identify immature-erythroblasts as the cells that generate additional erythroblasts in human-erythroid-massive-amplification cultures and autophagy as the leading cause of death limiting the final cellular output of these cultures.<br /> (Copyright © 2011 Elsevier Inc. All rights reserved.)
- Subjects :
- Anemia pathology
Apoptosis physiology
Blood Transfusion methods
Cell Differentiation
Cell Proliferation
Cell Separation
Cell Survival
Cells, Cultured
Coculture Techniques
Erythrocytes cytology
Glucocorticoids pharmacology
Growth Substances pharmacology
Humans
Immunophenotyping
Autophagy physiology
Cell Culture Techniques methods
Erythroblasts cytology
Erythropoiesis physiology
Hematopoietic Stem Cells cytology
Subjects
Details
- Language :
- English
- ISSN :
- 1096-0961
- Volume :
- 47
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Blood cells, molecules & diseases
- Publication Type :
- Academic Journal
- Accession number :
- 21775174
- Full Text :
- https://doi.org/10.1016/j.bcmd.2011.06.001