Application of mutant JAK2V617F for in vitro generation of red blood cells. Issue 4 (9th December 2015)

• Record Type: Journal Article
• Authors: Lee, Sun Ah; Kim, Ji Yeon; Choi, Yongwook; Kim, Yonggoo; Kim, Hyun Ok
• Is Part Of: Transfusion. Volume 56:Issue 4(2016:Apr.)
• Page Start: 837
• Page End: 843
• Publication Date: 2015-12-09
• Subjects: Hematology -- Periodicals; Blood -- Transfusion -- Periodicals; Blood Group Antigens -- Periodicals; Blood Preservation -- Periodicals; Blood Transfusion -- Periodicals; 615
• DOI: 10.1111/trf.13431
• Languages: English
• ISSNs: 0041-1132

Abstract

Abstract : 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.