An In Vitro Model of Human Red Blood Cell Production From Hematopoietic Progenitor Cells

• Published in: Blood Vol. 91; no. 8; pp. 2664 - 2671
• Main Authors: Malik, Punam, Fisher, Timothy C., Barsky, Lora L.W., Zeng, Licheng, Izadi, Parvin, Hiti, Alan L., Weinberg, Kenneth I., Coates, Thomas D., Meiselman, Herbert J., Kohn, Donald B.
• Format: Journal Article
• Language: English
• Published: Washington, DC Elsevier Inc 15-04-1998; The Americain Society of Hematology
• Subjects: Adult; Antigens, CD34; Biological and medical sciences; Cell Culture Techniques; Cell Differentiation; Cell differentiation, maturation, development, hematopoiesis; Cell physiology; Erythrocytes - cytology; Erythropoiesis; Flow Cytometry; Fundamental and applied biological sciences. Psychology; Genetic Markers; Hematopoietic Stem Cells - cytology; Humans; Molecular and cellular biology; Antigen; Erythropoiesis; Human; Genetic marker; Hematopoiesis; Stem cell; Biological model; Hematopoietic cell; Gene therapy; In vitro; Cell subpopulation; Cell surface
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V91.8.2664.2664_2664_2671

Hemoglobinopathies, such as β-thalassemias and sickle cell anemia (SCA), are among the most common inherited gene defects. Novel models of human erythropoiesis that result in terminally differentiated red blood cells (RBCs) would be able to address the pathophysiological abnormalities in erythrocytes in congenital RBC disorders and to test the potential of reversing these problems by gene therapy. We have developed an in vitro model of production of human RBCs from normal CD34+ hematopoietic progenitor cells, using recombinant growth factors to promote terminal RBC differentiation. Enucleated RBCs were then isolated to a pure population by flow cytometry in sufficient numbers for physiological studies. Morphologically, the RBCs derived in vitro ranged from early polylobulated forms, resembling normal reticulocytes to smooth biconcave discocytes. The hemoglobin pattern in the in vitro-derived RBCs mimicked the in vivo adult or postnatal pattern of β-globin production, with negligible γ-globin synthesis. To test the gene therapy potential using this model, CD34+ cells were genetically marked with a retroviral vector carrying a cell-surface reporter. Gene transfer into CD34+ cells followed by erythroid differentiation resulted in expression of the marker gene on the surface of the enucleated RBC progeny. This model of human erythropoiesis will allow studies on pathophysiology of congenital RBC disorders and test effective therapeutic strategies.