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Application of mutant JAK2 V617F for in vitro generation of red blood cells
Author(s) -
Lee Sun Ah,
Kim Ji Yeon,
Choi Yongwook,
Kim Yonggoo,
Kim Hyun Ok
Publication year - 2016
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/trf.13431
Subject(s) - haematopoiesis , erythropoiesis , cord blood , cd34 , stem cell , erythropoietin , biology , microbiology and biotechnology , cellular differentiation , interleukin 3 , erythropoietin receptor , immunology , medicine , antigen presenting cell , endocrinology , immune system , anemia , biochemistry , gene , t cell
BACKGROUND In vitro generation of red blood cells (RBCs) from hematopoietic stem cells (HSCs) has been reported, but the collection of 1 × 10 5 to 1 × 10 6 CD34+ cells present in cord and peripheral blood is too small for expansion to 1 × 10 12 cells in 1 unit of RBCs. We transduced JAK2 V617F gene, the most common mutation with polycythemia vera (PV), into cord blood–derived CD34+ cells. This PV model was expected to increase cell proliferation without the addition of erythropoietin (EPO) in early phase of differentiation. STUDY DESIGN AND METHODS Empty vector (control), wild‐type JAK2 (w JAK2 ), and mutant JAK2 V617F (m JAK2 ) were transduced into CD34+ cells using a lentivirus system. The CD34+ cells were then differentiated to the RBCs in a culture system. The cells were analyzed for cell number, differential count, and morphologic changes. Cultured RBCs were tested for oxygen equilibrium. RESULTS w JAK2‐ and m JAK2 ‐transduced cells showed higher proliferation capacity until Day 21 than control cells; interestingly, only m JAK2 ‐transduced cells were highly increased on Day 7 during EPO‐free culture. However, both w JAK2 ‐ and m JAK2 ‐tranduced cells had more delayed differentiation than control, but they had a higher portion of completely matured RBCs and orthochromatic erythroblasts. Furthermore, m JAK2 ‐tranduced cells showed more differentiation into RBCs than w JAK2 ‐transduced cells and they had a normal hemoglobin dissociation curve. CONCLUSION This is the first trial to use a PV erythropoiesis model for RBC differentiation from stem cells. The transduction of HSCs with m JAK2 increased their proliferation capacity in EPO‐free culture conditions. This model may also be useful for investigating the pathogenesis of PV.