Establishment of an erythroid cell line from primary CD36+ erythroid progenitor cells

• Published in: Experimental hematology 2010-11, Vol.38 (11), p.994-1005.e2
• Main Authors: Wong, Susan, Keyvanfar, Keyvan, Wan, Zhihong, Kajigaya, Sachiko, Young, Neal S, Zhi, Ning
• Format: Article
• Language: English
• Subjects: Advanced Basic Science; CD36 Antigens - genetics; CD36 Antigens - metabolism; Cell Differentiation - drug effects; Cell Line, Transformed; Cell Proliferation - drug effects; Cell Transformation, Viral; Cells, Cultured; Chromosome Banding; Colony-Forming Units Assay; Cytokines - pharmacology; Erythroid Cells - cytology; Erythroid Cells - metabolism; Erythroid Precursor Cells - cytology; Erythroid Precursor Cells - metabolism; Flow Cytometry; Gene Expression Profiling; Genetic Vectors - genetics; Hematology, Oncology and Palliative Medicine; Humans; Immunoblotting; Immunophenotyping; K562 Cells; Lentivirus - genetics; Oncogene Proteins, Viral - genetics; Oncogene Proteins, Viral - metabolism; Papillomavirus E7 Proteins - genetics; Papillomavirus E7 Proteins - metabolism; Repressor Proteins - genetics; Repressor Proteins - metabolism; Reverse Transcriptase Polymerase Chain Reaction; Spectral Karyotyping; Thrombopoietin - pharmacology
• ISSN: 0301-472X; 1873-2399
• DOI: 10.1016/j.exphem.2010.07.012

Objective Most continuous cell lines with erythroid characteristics are derived from patients with myelogenous leukemia or erythroleukemia. Among them, a few cell lines have been reported to be positive for CD36. We tried to establish a continuous erythroid cell line from the primary CD36+ erythroid progenitor cells (EPCs) by the lentivirus-mediated gene transduction system. Materials and Methods A lentiviral vector carrying SV40T , hTERT, or the human papillomavirus type 16 (HPV16) E6 and E7 ( E6 / E7 ) viral oncogenes, was introduced into CD36+ EPCs, singularly or combined. Transformed cells were characterized in terms of histology, phenotype, karyotype, and gene expression profile. Results The lentiviral vector carrying HPV16 E6/E7 genes successfully transformed CD36+ EPCs, creating a continuous cell line, CD36E. Immunophenotype analysis revealed that the CD36E cells had characteristics of erythroid progenitors, among which about 27% of the cell population produced hemoglobin. Colony-forming cell assay demonstrated that the CD36E cells were capable of forming erythroid colonies. Using cytokines or chemical agents, attempts were made to induce differentiation of the CD36E cells but were ineffective, indicating the irreversible erythroid lineage commitment of the cells. The gene expression profile of the CD36E cells displayed a marked difference from that of the CD36+ EPCs. Conclusions The continuous CD36E cell line is an erythroid progenitor cell line possessing the ability to produce hemoglobin. The CD36E cell line would be an excellent tool for applied research involving erythroid lineage cells and comparative studies with primary CD36+ EPCs.