1. K562 erythroleukemia line as a possible reticulocyte source to culture Plasmodium vivax and its surrogates
- Author
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Peter Milanov, Lars C Vermaat, Clemens H. M. Kocken, Erica M. Pasini, Jessica Thiel, Romy Kronstein-Wiedemann, Torsten Tonn, Onny Klop, and Claudia Ruhland
- Subjects
0301 basic medicine ,Cancer Research ,Reticulocytes ,Receptor expression ,Cellular differentiation ,Enucleation ,Population ,Receptors, Cell Surface ,Biology ,Article ,03 medical and health sciences ,0302 clinical medicine ,Reticulocyte ,hemic and lymphatic diseases ,parasitic diseases ,Genetics ,medicine ,Humans ,RNA, Neoplasm ,education ,Molecular Biology ,education.field_of_study ,Gene Expression Regulation, Leukemic ,Cell Differentiation ,Cell Biology ,Hematology ,Neoplasm Proteins ,Cell biology ,MicroRNAs ,Red blood cell ,030104 developmental biology ,medicine.anatomical_structure ,Cell culture ,030220 oncology & carcinogenesis ,Erythropoiesis ,Leukemia, Erythroblastic, Acute ,Duffy Blood-Group System ,K562 Cells ,Plasmodium vivax - Abstract
Highlights • miR-26a and miR-30a knockdowns promote differentiation in Fy-transduced K562 cell lines. • miR-26a and miR-30a knockdowns promote enucleation in Fy-transduced K562 cell lines. • Data denote an interplay in the mode of action of miR-26a and miR-30a in erythropoiesis. • Plasmodium cynomolgi and P. knowlesi invade, albeit inefficiently, Fy-transduced K562 cells., Establishing an in vitro “red blood cell matrix” that would allow uninterrupted access to a stable, homogeneous reticulocyte population would facilitate the establishment of continuous, long-term in vitro Plasmodium vivax blood stage cultures. In this study, we have explored the suitability of the erythroleukemia K562 cell line as a continuous source of such reticulocytes and have investigated regulatory factors behind the terminal differentiation (and enucleation, in particular) of this cell line that can be used to drive the reticulocyte production process. The Duffy blood group antigen receptor (Fy), essential for P. vivax invasion, was stably introduced into K562 cells by lentiviral gene transfer. miRNA-26a-5p and miRNA-30a-5p were downregulated to promote erythroid differentiation and enucleation, resulting in a tenfold increase in the production of reticulocytes after stimulation with an induction cocktail compared with controls. Our results suggest an interplay in the mechanisms of action of miRNA-26a-5p and miRNA-30a-5p, which makes it necessary to downregulate both miRNAs to achieve a stable enucleation rate and Fy receptor expression. In the context of establishing P. vivax-permissive, stable, and reproducible reticulocytes, a higher enucleation rate may be desirable, which may be achieved by the targeting of further regulatory mechanisms in Fy-K562 cells; promoting the shift in hemoglobin production from fetal to adult may also be necessary. Despite the fact that K562 erythroleukemia cell lines are of neoplastic origin, this cell line offers a versatile model system to research the regulatory mechanisms underlying erythropoiesis.
- Published
- 2020