Long-term ability for hematopoiesis of CD34(+)CD59(+) cells from patients with paroxysmal nocturnal hemoglobinuria: experiment with mice

To obtain the evidence of long-term hematopoiesis by normal clone stem cells from patients with paroxysmal nocturnal hemoglobinuria (PNH). 10 ml fresh bone marrow was collected from 2 PNH patients. Normal bone marrow was obtained from the ribs resected during operation of 2 patients without hematopathy. CD34(+)CD59(+) cells were isolated, purified, and expended. 12 total body irradiated SCID mice were divided into 2 equal group to undergo transplantation of the expanded CD34(+)CD59(+) cells from PNH patients (experiment group 1) or normal patients (control group 1). Four mice underwent transfusion of cell-free medium as blank control group 1. 30 and 60 days after the transplantation peripheral blood samples were collected and 90 days after the transplantation heart blood samples were collected. Spleens were taken out to prepare suspension of splenocytes and bacterium-free bone marrow cell suspension was prepared from the bilateral femurs. 90 days after the primary transplantation another 16 mice underwent tot